The effect of extracellular polymeric substances (EPS) on the meso-scale physical structure and hydraulic resistance of membrane biofilms during gravity driven membrane (GDM) filtration was ...investigated. Biofilms were developed on the surface of ultrafiltration membranes during dead-end filtration at ultra-low pressure (70 mbar). Biofilm EPS composition (total protein, polysaccharide and eDNA) was manipulated by growing biofilms under contrasting nutrient conditions. Nutrient conditions consisted of (i) a nutrient enriched condition with a nutrient ratio of 100:30:10 (C: N: P), (ii) a phosphorus limitation (C: N: P ratio: 100:30:0), and (iii) a nitrogen limitation (C: N: P ratio: 100:0:10). The structure of the biofilm was characterised at meso-scale using Optical Coherence Tomography (OCT). Biofilm composition was analysed with respect to total organic carbon, total cellular mass and extracellular concentrations of proteins, polysaccharides, and eDNA. 2D-confocal Raman mapping was used to characterise the functional group composition and micro-scale distribution of the biofilms EPS. Our study reveals that the composition of the EPS matrix can determine the meso-scale physical structure of membrane biofilms and in turn its hydraulic resistance. Biofilms grown under P limiting conditions were characterised by dense and homogeneous physical structures with high concentrations of polysaccharides and eDNA. Biofilm grown under nutrient enriched or N limiting conditions were characterised by heterogeneous physical structures with lower concentrations of polysaccharides and eDNA. For P limiting biofilms, 2D-confocal Raman microscopy revealed a homogeneous spatial distribution of anionic functional groups in homogeneous biofilm structures with higher polysaccharide and eDNA concentrations. This study links EPS composition, physical structure and hydraulic resistance of membrane biofilms, with practical relevance for the hydraulic performances of GDM ultrafiltration.
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•Hydraulic resistance of biofilms formed during GDM filtration was studied.•Influent composition determined EPS chemical composition.•Resulting EPS compositions were linked to distinct physical structures.•Hydraulic resistance was dependent on biofilm thickness and density.
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•Abundance, distribution and activity of AOB and anammox in aggregates were studied.•AOB were enriched in the floccular fraction, while anammox were enriched in granules.•Higher ...ratios of anammox/AOB were found in plants with hydrocyclones.•Substantial functional and population-level segregation in aggregates were found.
The purpose of this study was to determine the abundance, distribution and activity of aerobic ammonia-oxidizing bacteria (AOB) and anammox in size fractionated aggregates from full-scale suspended growth combined nitritation-anammox sidestream reactors. Plants with or without a cyclone device were also studied to assess a purported enrichment of anammox granules. Specific aerobic ammonium oxidation rates (p=0.01) and specific oxygen uptake rates (p=0.02) were significantly greater in flocs than in granules. AOB abundance measured using quantitative FISH was significantly higher in flocs than in granules (p=0.01). Conversely, anammox abundance was significantly greater in granules (p=0.03). The average ratio of anammox/AOB in systems employing hydrocyclone separation devices was 2.4, significantly higher (p=0.02) than the average ratio (0.5) in a system without a hydrocyclone. Our results demonstrate substantial functional and population-level segregation between floccular and granular fractions, and provide a key corroboration that cyclone separation devices can increase anammox levels in such systems.
This study aimed at evaluating the formation of aerobic granular sludge (AGS) for the treatment of real and low-strength municipal wastewater using a column sequencing batch reactor (SBR) operated in ...fill-draw mode (constant volume). The focus was on understanding how the wastewater upflow velocity (VWW) applied during the anaerobic feeding influenced the sludge properties and in turn the substrate conversion. Two different strategies were tested: (1) washing-out the flocs by imposing high wastewater upflow velocities (between 5.9 and 16 m h−1) during the anaerobic feeding (Approach #1) and (2) selective utilization of organic carbon during the anaerobic feeding (1 m h−1) combined with a selective sludge withdrawal (Approach #2). A column SBR of 190 L was operated in constant volume during 1500 days and fed with real and low-strength municipal wastewater. The formation of AGS with SVI30 of around 80 mL gTSS−1 was observed either at very low (1 m h−1) or at high VWW (16 m h−1). At 16 m h−1 the AGS was mainly composed of large and round granules (d > 0.63 mm) with a fluffy surface, while at 1 m h−1 the sludge was dominated by small granules (0.25 < d < 0.63 mm). The AGS contained a significant fraction of flocs during the whole operational period. A considerable and continuous washout of biomass occurred at VWW higher than 5.9 m h−1 (Approach #1) due to the lower settling velocity of the AGS fed with municipal wastewater. The low sludge retention observed at VWW higher than 5.9 m h−1 deteriorated the substrate conversion and in turn the effluent quality. High solid concentrations (and thus solid retention time) were maintained during Approach #2 (VWW of 1 m h−1), which resulted in an excellent effluent quality. The study demonstrated that the formation of AGS is possible during the treatment of real and low-strength municipal wastewater in a SBR operated at constant volume. Low wastewater upflow velocities should be applied during the anaerobic feeding phase in order to ensure enough biomass retention and efficient substrate removal.
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•Formation of aerobic granules with low-strength municipal wastewater was studied.•Different wastewater upflow velocities were tested during the anaerobic feeding.•Successful granule formation at low (1 m h−1) or high (16 m h−1) VWW.•High substrate conversion at VWW at 1 m h−1 due to a sufficient sludge retention.•No simultaneous nitrification-denitrification was observed in our AGS reactor.
The implementation of autotrophic anaerobic ammonium oxidation processes for the removal of nitrogen from municipal wastewater (known as “mainstream anammox”) bears the potential to bring wastewater ...treatment plants close to energy autarky. The aim of the present work was to assess the long-term stability of partial nitritation/anammox (PN/A) processes operating at low temperatures and their reliability in meeting nitrogen concentrations in the range of typical discharge limits below 2 mgNH4-N·L−1 and 10 mgNtot·L−1. Two main 12-L sequencing batch reactors were operated in parallel for PN/A on aerobically pre-treated municipal wastewater (21 ± 5 mgNH4-N·L−1 and residual 69 ± 19 mgCODtot·L−1) for more than one year, including over 5 months at 15 °C. The two systems consisted of a moving bed biofilm reactor (MBBR) and a hybrid MBBR (H-MBBR) with flocculent biomass. Operation at limiting oxygen concentrations (0.15–0.18 mgO2·L−1) allowed stable suppression of the activity of nitrite-oxidizing bacteria at 15 °C with a production of nitrate over ammonium consumed as low as 16% in the MBBR. Promising nitrogen removal rates of 20–40 mgN·L−1·d−1 were maintained at hydraulic retention times of 14 h. Stable ammonium and total nitrogen removal efficiencies over 90% and 70% respectively were achieved. Both reactors reached average concentrations of total nitrogen below 10 mgN·L−1 in their effluents, even down to 6 mgN·L−1 for the MBBR, with an ammonium concentration of 2 mgN·L−1 (set as operational threshold to stop aeration). Furthermore, the two PN/A systems performed almost identically with respect to the biological removal of organic micropollutants and, importantly, to a similar extent as conventional treatments. A sudden temperature drop to 11 °C resulted in significant suppression of anammox activity, although this was rapidly recovered after the temperature was increased back to 15 °C. Analyses of 16S rRNA gene-targeted amplicon sequencing revealed that the anammox guild of the bacterial communities of the two systems was composed of the genus “Candidatus Brocadia”. The potential of PN/A systems to compete with conventional treatments for biological nutrients removal both in terms of removal rates and overall effluent quality was proven.
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•Stable partial nitritation/anammox of aerobically pre-treated municipal wastewater at 15 °C.•Effluent ammonia and total nitrogen concentrations meet current discharge limits.•Successful NOB activity suppression at low dissolved oxygen concentrations (0.15–0.18 mgO2/L).•Micropollutants removal comparable to conventional biological treatments.•Reversible but dramatic anammox activity suppression during prolonged operation at 11 °C.
The control of nitrite-oxidizing bacteria (NOB) challenges the implementation of partial nitritation and anammox (PN/A) processes under mainstream conditions. The aim of the present study was to ...understand how operating conditions impact microbial competition and the control of NOB in hybrid PN/A systems, where biofilm and flocs coexist. A hybrid PN/A moving-bed biofilm reactor (MBBR; also referred to as integrated fixed film activated sludge or IFAS) was operated at 15 °C on aerobically pre-treated municipal wastewater (23 mgNH4-N L−1). Ammonium-oxidizing bacteria (AOB) and NOB were enriched primarily in the flocs, and anammox bacteria (AMX) in the biofilm. After decreasing the dissolved oxygen concentration (DO) from 1.2 to 0.17 mgO2 L−1 - with all other operating conditions unchanged - washout of NOB from the flocs was observed. The activity of the minor NOB fraction remaining in the biofilm was suppressed at low DO. As a result, low effluent NO3− concentrations (0.5 mgN L−1) were consistently achieved at aerobic nitrogen removal rates (80 mgN L−1 d−1) comparable to those of conventional treatment plants. A simple dynamic mathematical model, assuming perfect biomass segregation with AOB and NOB in the flocs and AMX in the biofilm, was able to qualitatively reproduce the selective washout of NOB from the flocs in response to the decrease in DO-setpoint. Similarly, numerical simulations indicated that flocs removal is an effective operational strategy to achieve the selective washout of NOB. The direct competition for NO2− between NOB and AMX - the latter retained in the biofilm and acting as a “NO2-sink” - was identified by the model as key mechanism leading to a difference in the actual growth rates of AOB and NOB (i.e., μNOB < μAOB in flocs) and allowing for the selective NOB washout over a broad range of simulated sludge retention times (SRT = 6.8–24.5 d). Experimental results and model predictions demonstrate the increased operational flexibility, in terms of variables that can be easily controlled by operators, offered by hybrid systems as compared to solely biofilm systems for the control of NOB in mainstream PN/A applications.
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•Hybrid PN/A systems provide increased operational flexibility for NOB control•AOB and NOB enrich primarily in the flocs, and AMX in the biofilm (“NO2-sink”)•AMX use NO2− allowing to differentiate AOB and NOB growth rates•A decrease in DO or an increase in floc removal leads to selective NOB washout from flocs•The activity of the minor NOB fraction in the biofilm is suppressed at limiting DO
Direct treatment of municipal wastewater (MWW) based on anaerobic ammonium oxidizing (anammox) bacteria holds promise to turn the energy balance of wastewater treatment neutral or even positive. ...Currently, anammox processes are successfully implemented at full scale for the treatment of high-strength wastewaters, whereas the possibility of their mainstream application still needs to be confirmed. In this study, the growth of anammox organisms on aerobically pre-treated municipal wastewater (MWWpre-treated), amended with nitrite, was proven in three parallel reactors. The reactors were operated at total N concentrations in the range 5–20 mgN∙L−1, as expected for MWW. Anammox activities up to 465 mgN∙L−1∙d−1 were reached at 29 °C, with minimum doubling times of 18 d. Lowering the temperature to 12.5 °C resulted in a marked decrease in activity to 46 mgN∙L−1∙d−1 (79 days doubling time), still in a reasonable range for autotrophic nitrogen removal from MWW. During the experiment, the biomass evolved from a suspended growth inoculum to a hybrid system with suspended flocs and wall-attached biofilm. At the same time, MWWpre-treated had a direct impact on process performance. Changing the influent from synthetic medium to MWWpre-treated resulted in a two-month delay in net anammox growth and a two to three-fold increase in the estimated doubling times of the anammox organisms. Interestingly, anammox remained the primary nitrogen consumption route, and high-throughput 16S rRNA gene-targeted amplicon sequencing analyses revealed that the shift in performance was not associated with a shift in dominant anammox bacteria (“Candidatus Brocadia fulgida”). Furthermore, only limited heterotrophic denitrification was observed in the presence of easily biodegradable organics (acetate, glucose). The observed delays in net anammox growth were thus ascribed to the acclimatization of the initial anammox population or/and the development of a side population beneficial for them. Additionally, by combining microautoradiography and fluorescence in situ hybridization it was confirmed that the anammox organisms involved in the process did not directly incorporate or store the amended acetate and glucose. In conclusion, these investigations strongly support the feasibility of MWW treatment via anammox.
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•Anammox guild grows on aerobically pre-treated municipal wastewater (MWW) amended with NO2−.•Anammox activity at 12.5 °C was comparable to conventional N-removal treatments.•Anammox growth was 2 to 3 times slower on pre-treated compared to synthetic MWW.•“Ca. Brocadia fulgida” was the dominant anammox strain growing on pre-treated MWW.•“Ca. Brocadia” affiliates did not directly accumulate organics (acetate, glucose).
This study evaluates the effect of the presence of biofilms on membrane surfaces on the quality of permeate produced during Gravity-driven membrane ultrafiltration. GDM ultrafiltration is applied to ...the decentralized production of drinking water. A second objective was to evaluate to what extent permeate quality is enhanced by pre-treating feed-water (using a packed bed biofilm reactor or a slow sand filter). The influence of the ageing of the biofilm on the permeate quality was evaluated and compared to the effect of virgin membranes. Permeate quality was evaluated in terms of Assimilable Organic Carbon (AOC) content and dissolved organic carbon fractions (e.g. biopolymers). Our results indicate that virgin ultrafiltration membrane remove a small fraction of the AOC and biopolymers (rejection <10%). The presence of a young and thin biofilm on the surface of the ultrafiltration membranes increases the permeate quality due to the degradation of AOC (>80%). However, over long-term the hydrolysis of the organic matter that accumulated on membrane surfaces increases the AOC content of the permeate, thus deteriorating the permeate quality. Pre-treatment of the feed-water help to control the biofilm accumulation and thus to limit the deterioration of the permeate quality. Permeate flux stabilised at average values of 7.5-8.9 L m(-2) h(-1). But the presence of pre-treatment helped to increase permeate flux (+12 and 19%, with the packed bed biofilm reactor and with the slow sand filter, respectively). Overall our study demonstrates that tolerating the presence of biofilm on membrane surface has a beneficial effect on the quality of permeate even if its quantity is decreased.
This study investigates the impact of predation by eukaryotes on the development of specific biofilm structures in gravity-driven dead-end ultrafiltration systems. Filtration systems were operated ...under ultra-low pressure conditions (65 mbar) without the control of biofilm formation. Three different levels of predation were evaluated: (1) inhibition of eukaryotic organisms, (2) addition of cultured protozoa (Tetrahymena pyriformis), and (3) no modification of microbial community as a control. The system performance was evaluated based on permeate flux and structures of the biofilm. It was found that predation had a significant influence on both the total amount and also the structure of the biofilm. An open and heterogeneous structure developed in systems with predation whereas a flat, compact, and thick structure that homogeneously covered the membrane surface developed in absence of predation. Permeate flux was correlated with the structure of the biofilm with increased fluxes for smaller membrane coverage. Permeate fluxes in the presence or absence of the predators was 10 and 5 L m−2 h−1, respectively. It was concluded that eukaryotic predation is a key factor influencing the performance of gravity-driven ultrafiltration systems.
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► Predation affects biofilms developed during gravity-driven dead-end ultrafiltration. ► Predation resulted in more open and heterogeneous biofilm structures. ► Open and heterogeneous structures induced higher permeate fluxes.
This study investigates the functional correspondence between porescale hydrodynamics, mass transfer, pore structure and biofilm morphology during progressive biofilm colonization of a porous medium. ...Hydrodynamics and the structure of both the porous medium and the biofilm are experimentally measured with 3D particle tracking velocimetry and micro X-ray Computed Tomography, respectively. The analysis focuses on data obtained in a clean porous medium after 36 h of biofilm growth. Registration of the particle tracking and X-ray data sets allows to delineate the interplay between porous medium geometry, hydrodynamic and mass transfer processes on the morphology of the developing biofilm. A local analysis revealed wide distributions of wall shear stresses and concentration boundary layer thicknesses. The spatial distribution of the biofilm patches uncovered that the wall shear stresses controlled the biofilm development. Neither external nor internal mass transfer limitations were noticeable in the considered system, consistent with the excess supply of nutrient and electron acceptors. The wall shear stress remained constant in the vicinity of the biofilm but increased substantially elsewhere.
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