Lab-scale Moving Bed Biofilm Reactor Research Articles

The Biodegradation of 4-Chlorophenol in a Moving Bed Biofilm Reactor Using Response Surface Methodology: Effect of Biogenic Substrate and Kinetic Evaluation.

4-Chlorophenol (4-CP) is a persistent organic pollutant commonly found in petrochemical effluents. It causes toxic, carcinogenic and mutagenic effects on human beings and aquatic lives. Therefore, an environmentally benign and cost-effective approach is needed against such pollutants. In this direction, the chlorophenol degrading bacterial consortium consisting of Bacillus flexus GS1 IIT (BHU) and Bacillus cereus GS2 IIT (BHU) was isolated from a refinery site. A composite biocarrier namely polypropylene-polyurethane foam (PP-PUF) was developed for bacterial cells immobilization purpose. A lab-scale moving bed biofilm reactor (MBBR) packed with Bacillus sp. immobilized PP-PUF biocarrier was employed to analyse the effect of peptone on biodegradation of 4-CP. The statistical tool, i.e. response surface methodology (RSM), was used to optimize the process variables (4-CP concentration, peptone concentration and hydraulic retention time). The higher values of peptone concentration and hydraulic retention time were found to be favourable for maximum removal of 4-CP. At the optimized process conditions, the maximum removals of 4-CP and chemical oxygen demand (COD) were obtained to be 91.07 and 75.29%, respectively. In addition, three kinetic models, i.e. second-order, Monod and modified Stover-Kincannon models, were employed to investigate the behaviour of MBBR during 4-CP biodegradation. The high regression coefficients obtained by the second-order and modified Stover-Kincannon models showed better accuracy for estimating substrate degradation kinetics. The phytotoxicity study supported that the Vigna radiata seeds germinated in treated wastewater showed higher growth (i.e. radicle and plumule) than the untreated wastewater.

Effect of mixing intensity on biodegradation of phenol in a moving bed biofilm reactor: Process optimization and external mass transfer study

In this work, an effort has been made to design the process variables and to analyse the impact of mixing intensity on mass transfer diffusion in a moving bed biofilm reactor (MBBR). A lab-scale MBBR, filled with Bacillus cereus GS2 IIT (BHU) immobilized-polyethylene biocarriers, was employed to optimize the process variables, including mixing intensity (60–140 rpm), phenol concentration (50–200 mg/L), and hydraulic retention time (HRT) (4–24 h) using response surface methodology. The optimum phenol removal of 87.64 % was found at 100 rpm of mixing intensity, 200 mg/L of phenol concentration, and 24 h of HRT. The higher mixing intensity improved the substrate diffusion between the liquid phase and the surface of the biofilm. The external mass transfer coefficients were found in the range of 1.431 × 10-5-1.845 × 10-5 m/s. Moreover, the detection of catechol and 2-hydroxymuconic semialdehyde revealed that the Bacillus sp. followed the meta-cleavage pathway during the biodegradation of phenol.

Novel intertidal wetland sediment-inoculated moving bed biofilm reactor treating high-salinity wastewater: Metagenomic sequencing revealing key functional microorganisms

In this study, two lab-scale moving bed biofilm reactors (MBBR), seeded with intertidal wetland sediment (IWS) and activated sludge (AS), were constructed to compare their performances in treating high-salinity (3%) wastewater. Under a wide range of influent TOC (178–620 mg/L) and NH4+-N (25–100 mg/L), both the MBBRs (Riws and Ras) exhibited excellent TOC removal efficiencies of >95%. Regarding nitrogen reduction, Riws exhibited a significantly superior TN removal efficiency of 90.2 ± 1.8% than that of Ras (76.8 ± 2.9%). A correlation analysis was innovatively conducted comparing the results between metagenomic sequencing and DNA pyrosequencing, and positive linear relationships were found with R2 values of 0.763–0.945. Meanwhile, for illustration of different TN removal performance, nitrogen metabolic pathways were also assessed. Moreover, a list of functional oxidases (EC: 1.13.11.1, EC: 1.13.11.2, EC: 1.13.11.24, EC: 1.13.12.16, EC: 1.4.3.4, EC: 1.16.3.3, EC: 1.14.14.28) was found in IWS, revealing its potential in degradation of recalcitrant organics.

Achieving Partial Nitritation in Anammox Start-Up Environment

Removing ammonium via the partial nitritation anammox (PNA) process has been widely applied because of its cost and energy effectiveness. However, the first stage of PNA, partial nitritation, is hard to implement practically due to the challenging suppression of nitrate oxidizing bacteria (NOB) and should be achieved in the anammox environment to extend it to one stage PNA. Hence, this article evaluates different techniques, such as the combination of low dissolve oxygen (DO) and high free ammonia (FA), and the intermittent aeration cycle to achieve partial nitritation in an anammox start-up environment. For this purpose, a 10.5 L lab-scale moving bed biofilm reactor was set up and fed with synthetic wastewater and the transformation of influent ammonium into nitrate and nitrite was measured. The results showed that, despite applying low DO and higher free ammonia than the inhibition range of NOB, the nitrate production rate (NPR) was consistently higher than the nitrite accumulation rate (NAR), signifying no sufficient NOB suppression, partial nitritation under continuous aeration and up to a 0.27 gN/m2·d surface ammonium loading rate (SALR). Higher SALR than 0.27 gN/m2·d could result in partial nitritation since nitrogen compounds transformation was closer to partial nitritation when the reactor was subjected to 0.27 gN/m2·d rather than 0.14 gN/m2·d. Lifting up the SALR, on the other hand, results in a bad anammox environment and cannot prolong it to one-stage PNA. An intermittent aeration cycle with four different cycle lengths sets, obtained by monitoring nitrogen compound transformation, was, therefore, applied to the reactor. The relatively shorter aerobic length of 10 min ON and 30 OFF intermittent aeration cycle with 0.5 mg/L aerated DO was successful in achieving the partial nitritation with NPR, NAR, and ammonium removal efficiency (ARE) values of 17%, 78%, and 37%, respectively, showing that shorter aerated length suppresses NOB to a high degree due to less available time for NOB after oxygen starvation.

Impact of Saddle-Chips biocarrier on treating mariculture wastewater by moving bed biofilm reactor (MBBR): Mechanism and kinetic study

Packing carrier is a key factor for moving bed biofilm reactors (MBBRs) treating ammonia polluted wastewater such as in mariculture aquaculture but lacks sufficient research. In this study, a new type of biocarrier, Saddle-Chips (SC), was applied for treating mariculture wastewater (30‰ salinity) and compared with commonly used Kaldnes-5 (K5-biocarrier), using lab-scale MBBRs. The effects of ammonia concentrations, hydraulic retention times (HRTs), filling ratios (FRs), and ammonia shock-loading on MBBRs’ performances were examined. In the results, the new SC-biocarrier showed a slightly higher NH4+−N removal efficiency (NRE) than K5 (p > 0.05). Higher HRTs (24 and 12 h) yielded higher nutrients removal efficiency despite low biomass productivities initially. Moreover, 60% FR showed higher nutrients removal due to higher biomass than 30% FR (p < 0.05). Meanwhile, biopolymers released from the SC-biocarrier and ammonia shock-loading resulted in significantly (p < 0.05) higher Nitrospira abundance in MBBRs containing SC-biocarriers. Biofilm characterization through Fourier Transform Infrared (FTIR) Spectroscopy and Scanning Electron Microscope (SEM) revealed proteins, carbohydrates, lipids, RNA, and DNA traces in the biofilms involved. The Grau’s 2nd-order kinetics employed fitted well to the obtained results, predicting effluent NH4+−N concentrations and HRTs with high precision (R2 > 0.98). Overall, the nitrogenous pollutants’ high removal efficacy, biopolymer release capability, and excellent hydrodynamics revealed by the SC-biocarrier highlight its large-scale suitability in treating mariculture wastewater and other wastewater types.

Enhancement of nitrite/ammonia removal from saline recirculating aquaculture wastewater system using moving bed bioreactor

Innovative removal of toxic nitrite and ammonia is one of the key approaches for efficient aquaculture biotechnology. This study suggests a fast removal technique of high-levels of nitrite and ammonia using recirculating aquaculture wastewater at 30‰ salinity. Four lab-scale moving bed biofilm reactors (MBBRs; R1, R2, R3, and R4) were operated in parallel for 56 days with different combinations of mature and fresh biofilms on biocarriers at 30% filling ratio. Nitrogen removal efficiency, biofilm physicochemical properties, and microbial analyses were conducted using PCR and sequencing techniques at the region of the 16S ribosomal RNA. The results showed that R4 (using mature biofilms) recorded the highest nitrite and ammonia removal (99.6% and 95.3%, respectively) compared to the control, R1 (94.1% and 89.4%). Meanwhile, the removal rate of ammonia in R4 was significantly the fastest compared to other reactors. Partial Least Squares-Discriminant Multivariate Analysis (PLS-DA) model showed a good prediction with very high accuracy and prediction (Q2 > 0.8) for the first three principal components pertaining to nitrite and ammonia removal. Planktosalinus genus was the most abundant in R4, while proteobacteria phylum was the dominant, followed by Marinobacter and Nitrospina in all reactors. Overall, the use of mature biofilms in MBBR was proven to increase bacterial diversity, enhancing nitrite and ammonia removal efficiency.

Biodegradation of organophosphorus pesticides in moving bed biofilm reactors: Analysis of microbial community and biodegradation pathways

We investigated the performance of a lab-scale moving bed biofilm reactor (MBBR) with respect to general bioconversion processes and biotransformation of two commonly used organophosphorus pesticides, Chlorpyrifos (CHL) and Malathion (MAL). The reactor was operated for 300 days under different organic loads by changing hydraulic retention time (HRT). The decrease in organic load resulted in the formation of a thinner biofilm and the growth of more biomass in the bulk, which greatly shifted bioconversion processes. The low organic loading supported more nitrification in the reactor, but an opposite trend was observed for denitrification, which was enhanced at higher organic loading where the formation of anoxic zones in the thick biofilm was favored. 70% and 55% removal corresponding to 210 and 165 µg/m2/d occurred for MAL and CHL, respectively, at an HRT of 3 h and progressively increased with higher HRTs. Phylogenetic analysis revealed a shift in composition and abundance of taxa throughout the reactor operation where lower loading rate supported the growth of a more diverse and evenly distributed community. The analysis also highlighted the dominance of heterotrophic communities such as Flavobacterium and Acinetobacter johnsonii, which could be involved in the biotransformation of CHL and MAL through co-metabolism.

Anammox bacteria enrichment and denitrification in moving bed biofilm reactors packed with different buoyant carriers: Performances and mechanisms

Anaerobic ammonium oxidation (anammox) is recognized as the most cost-effective process for nitrogen removal from wastewater. In this study, effects of polyethylene plastics, nonwoven fabric, granular activated carbon (GAC) and polyurethane sponge as buoyant carriers were evaluated in lab-scale moving bed biofilm reactors (MBBRs). The overall performance of MBBRs with four types of carriers from priority to inferiority was noticed as, GAC, nonwoven fabrics, polyurethane sponge and polyethylene plastics under the same packing ratio of 20 v% and an average carrier size of 4 × 4 × 4 mm. The hydrophobic surface of GAC could selectively adsorb hydrophobic protein and favor anammox bacteria attachment, which contributed to achieving a total nitrogen removal rate of 0.40 kg-N/(m3·d) in 60 days. In conclusion, our results provide compelling evidence for achieving effective anammox process in an MBBR with GAC carriers and would benefit towards accomplishing a stable partial nitritation-anammox process in the future.

Efficient nitrous oxide recovery from incineration leachate by a nosZ-deficient strain of Pseudomonas aeruginosa

In this study, nitrous oxide was recovered from a lab-scale moving-bed biofilm reactor (MBBR) treating partial nitrification-treated leachate supplemented with a nosZ-deficient strain of Pseudomonas aeruginosa. Batch culture tests with the nosZ-deficient strain determined that the threshold for free nitrous acid (FNA) inhibition was 0.016 mg/L and that FNA concentrations above this threshold severely inhibited denitrification and transcription of genes from the dissimilatory nitrate reduction pathway (narG, nirS, and norB). High nitrite removal and N2O conversion efficiencies (>95%) were achieved with long-term operation of this MBBR. N2O accounted for the majority of biogas (80%) produced when the MBBR was fed partial nitrification-treated leachate with high nitrite concentrations and the drainage ratio was adjusted to 30%. Bacterial community analysis revealed that the nosZ-deficient Pseudomonas strain remained metabolically active and was primarily responsible for denitrification processes in the reactor. This study presents a promising method for N2O recovery from incineration leachate.

Impacts of typical pharmaceuticals and personal care products on the performance and microbial community of a sponge-based moving bed biofilm reactor

Four lab-scale moving bed biofilm reactors (MBBRs) were built to treat simulated wastewater containing typical pharmaceuticals and personal care products (PPCPs). The efficiency in removing different PPCPs at different concentrations (1, 2 and 5 mg/L) and their effects on the performance of MBBRs were investigated. Results showed that the average removal efficiencies of sulfadiazine, ibuprofen and carbamazepine were 61.1 ± 8.8%, 74.9 ± 8.8% and 28.3 ± 7.4%, respectively. Compared to the reactor without PPCPs, the total nitrogen (TN) removal efficiency of the reactors containing sulfadiazine, ibuprofen and carbamazepine declined by 21%, 30% and 42%, respectively. Based on the microbial community analysis, increasing the PPCPs concentration within a certain range (<2 mg/L) could stimulate microbial growth and increase microbial diversity yet the diversity reduced when the concentration (5 mg/L) exceeded the tolerance of microorganisms. Furthermore the presence and degradation of different PPCPs resulted in a different kind of microbial community structure in the MBBRs.

Elucidating the biofilm properties and biokinetics of a sulfur-oxidizing moving-bed biofilm for mainstream nitrogen removal

The sulfide-oxidizing autotrophic denitrification (SOAD) process offers a feasible alternative to mainstream heterotrophic denitrification in treating domestic sewage with insufficient organics. Previously SOAD has been successfully applied in a moving-bed biofilm reactor (MBBR). However, the biofilm properties and biokinetics are still not thoroughly understood. The present study was therefore designed to investigate these features of sulfur-oxidizing biofilms (SOBfs) cultivated in a lab-scale MBBR under stable operation for over a year. The biofilms developed were 160 μm thick, had an uneven and porous surface on which elemental sulfur (S0) accumulated, and the SOB biomass was highly diverse. The bioprocess kinetics were evaluated through 12 batch experiments. The results were interpreted by adopting a two-step sulfide oxidation model (sulfide→S0 and S0→ sulfate) with all specific rates having a linear regression coefficient of R2 > 0.9. Moreover, the inhibitory kinetic analysis revealed that 1) the maximum treatment capacity (about 480 mg S/(m2·h) and 80 mg N/(m2·h)) was observed at low sulfide level (40 mg S/L), while higher sulfide level (60-150 mg S/L) showed increasing inhibition on the oxidation of both sulfide and sulfur and denitrification. 2) The denitritation activity decreased by up to 43% when free nitrous acid reached a maximum of 8.6 μg N/L, whereas the oxidation of sulfide and sulfur did not have any significant effect. Interestingly, two physiologically diverse SOB groups were found in this special biofilm. The mechanisms of the cooperation and competition for electron donors and acceptors between these two SOB clades are proposed. The results of this study greatly enhance our understanding of the design and optimization of SOAD-MBBR for mainstream nitrogen removal.

Denitrification and microbial community in MBBR using A. donax as carbon source and biofilm carriers for reverse osmosis concentrate treatment

In this study, raw Arundo donax (A. donax) pieces were applied as carbon source and biofilm carriers for denitrification in a lab-scale moving bed biofilm reactor (MBBR) for the treatment of reverse osmosis concentrate gathered from local wastewater reuse plant. At stable phase (about 60 days), efficient denitrification performance was obtained with 73.2% ± 19.5% NO3−-N average removal and 8.10 ± 3.45 g N/(m3·day) NO3−-N average volumetric removal rate. Mass balance analysis showed that 4.84 g A. donax was required to remove 1 g TN. Quantitative real-time PCR analysis results showed that the copy numbers of 16S r-RNA, narG, nirS, nosZ and anammox gene of carrier biofilm and suspended activated sludge in the declination phase (BF2 and AS2) were lower than those of samples in the stable phase (BF1 and AS1), and relatively higher copy numbers of nirS and nirK genes with lower abundance of narG and nosZ genes were observed. High-throughput sequencing analysis was conducted for BF2 and AS2, and similar dominant phyla and classes with different abundance were obtained. The class Gammaproteobacteria affiliated with the phylum Proteobacteria was the most dominant microbial community in both BF2 (52.6%) and AS2 (41.7%). The PICRUSt prediction results indicated that 33 predictive specific genes were related to denitrification process, and the relative abundance of 18 predictive specific genes in BF2 were higher than those in AS2.

Performance evaluation of a lab-scale moving bed biofilm reactor (MBBR) using polyethylene as support material in the treatment of wastewater contaminated with terephthalic acid

Untreated terephthalic acid (TPA) wastewaters with high organic loads will cause severe environmental pollution problems. In this study, a lab-scale moving bed biofilm reactor, where biomass of Delftia sp. WL-3 is attached to polypropylene carrier elements, has been tested for TPA bioremediation at 25–27 °C. The system achieved stable operation after a short 15-day start-up period. During the operation period of 65 days, stable chemical oxygen demand (COD) and TPA removal efficiencies of 68% and 76% were maintained with an organic load rate (OLR) and hydraulic retention time of 2.5 kg COD·(m3·d)−1 and 24 h, respectively. In addition, the Scanning Electron Microscope (SEM) showed that high-densities of WL-3 biomass accumulated on the surface of the carrier and formed a rich biofilm, indicating polypropylene carrier can improve the degradation efficiency. On the contrary, the biodegradation ability of stain WL-3 without the polypropylene carrier declined significantly with removal efficiencies of 10% and 15% for COD and TPA. Furthermore, the system exhibited excellent robustness to different OLR and influent matrix ratios, indicating its potential for applications in the treatment of TPA-containment wastewater in the field.

Effect of adding low-concentration of rhamnolipid on reactor performances and microbial community evolution in MBBRs for low C/N ratio and antibiotic wastewater treatment

This study aims to explore the potential of low-concentration of rhamnolipid in efficient treatment of wastewater with poor biodegradability. Six lab-scale moving bed biofilm reactors (MBBRs) were applied to investigate the effect of rhamnolipid concentration (0, 20, 50 mg/L) on pollutants removal, biomass accumulation, microbial morphology and community evolution in synthetic low C/N ratio (3:1) and antibiotic (50 μg/L tetracycline) wastewater. 20 mg/L rhamnolipid treated groups exhibited significant increase (p < 0.05) of chemical oxygen demand (COD) removal and volatile solid (VS) content in both synthetic wastewater. Hydrogenophaga and Aeromonas were dominant in all reactors in which Aeromonas was positively correlated with the removal of COD and ammonia nitrogen (NH4+-N). Besides, Methyloversatilis became dominant only in 20 mg/L rhamnolipid treated groups and was positively correlated with VS. This study provides a novel and feasible strategy for treating poorly biodegradable wastewater by biofilm process with moderate amount of rhamnolipid.

Compact secondary treatment train combining a lab-scale moving bed biofilm reactor and enhanced flotation processes

High-rate wastewater processes are receiving a renewed interest to obtain energy positive/efficient water resource recovery facilities. An innovative treatment train combining a high-rate moving bed biofilm reactor (HR-MBBR) with an enhanced flotation process was studied. The two objectives of this work were 1) to maximize the conversion of soluble organics to particulate matter in an HR-MBBR and 2) to maximize the particulate matter recovery from the HR-MBBR effluent by green chemicals to enhance biogas production by anaerobic digestion. To achieve these objectives, lab-scale MBBRs fed with synthetic soluble wastewater were operated at organic loading rates (OLRs) between 4 and 34 kg COD m−3 reactor d−1 corresponding to hydraulic retention times (HRTs) between 6 and 54 min.Colloidal and soluble chemical oxygen demand (COD) removal efficiency in the HR-MBBR increased with HRT to reach a plateau of 85% at an HRT longer than 27 min. Carrier clogging observed at an OLR higher than 16 kg COD m−3 d−1 (HRT < 13 min) resulted in about 23% loss in colloidal and soluble COD removal efficiency. Thus, the recommended parameters were between 22 and 37 min and between 6 and 10 kg COD m−3 d−1 for the HRT and the OLR, respectively, to maximize the conversion of soluble organics to particulate matter.Total suspended solids (TSS) recovery of 58–85% and 90–97% were achieved by enhanced flotation using green and unbiodegradable chemicals, respectively, corresponding to a TSS effluent concentration below 14 and 7 mg TSS/L. Among the synthetic polymers tested, a high molecular weight and low charge density cationic polyacrylamide was found to give the best results with less than 2 mg TSS/L in the clarified effluent (97% TSS recovery). Green chemicals, although performing slightly less for solids separation than unbiodegradable chemicals, achieved a mean TSS concentration of 10 ± 3 mg/L in the clarified effluent.

An experimental model of COD abatement in MBBR based on biofilm growth dynamic and on substrates' removal kinetics

ABSTRACTIn this study, the performance of a lab-scale Moving Bed Biofilm Reactor (MBBR) under different operating conditions was analysed. Moreover, the dependence of the reaction rates both from the concentration and biodegradability of substrates and from the biofilm surface density, by means of several batch kinetic tests, was investigated. The reactor controls exhibited an increasing COD (Chemical Oxygen Demand) removal, reaching maximum yields (close to 90%) for influent loadings of up to12.5 gCOD/m2d. From this value, the pilot plant performance decreased to yields of only about 55% for influent loadings greater than 16 gCOD/m2d. In response to the influent loading increase, the biofilm surface density exhibited a logistic growing trend until reaching a maximum amount of total attached solids of about 9.5 g/m2. The kinetic test results indicated that the COD removal rates for rapidly biodegradable, rapidly hydrolysable and slowly biodegradable substrates were not affected by the organic matter concentrations. Instead, first-order kinetics were detected with respect to biofilm surface density. The experimental results permitted the formulation of a mathematical model to predict the MBBR organic matter removal efficiency. The validity of the model was successfully tested in the lab-scale plant.

Performance and modeling of a moving bed biofilm process: nickel and chromium heavy metal removal from industrial wastewater

The process of a lab-scale moving bed biofilm reactor (MBBR) using simulated sugar-manufacturing wastewater as feed was investigated.

Investigating biofilm structure developing on carriers from lab-scale moving bed biofilm reactors based on light microscopy and optical coherence tomography

This study focused on characterizing the structure of biofilms developed on carriers used in lab-scale moving bed biofilm reactors. Both light microscopy (2D) and optical coherence tomography (OCT) were employed to track the biofilm development on carriers of different geometry and under different aeration rates. Biofilm structure was further characterized with respect to average biofilm thickness, biofilm growth velocity, biomass volume, compartment filling degree, surface area, etc. The results showed that carriers with a smaller compartment size stimulated a quick establishment of biofilms. Low aeration rates favored fast development of biofilms. Comparison between the results derived from 2D and 3D images revealed comparable results with respect to average biofilm thickness and compartment filling degree before the carrier compartments were fully willed with biomass. However, 3D imaging with OCT was capable of visualizing and quantifying the heterogeneous structure of biofilms, which cannot be achieved using 2D imaging.

Determining the flow regime in a biofilm carrier by means of magnetic resonance imaging.

Biofilms on cylindrical carrier material originating from a lab-scale moving bed biofilm reactor (MBBR) were investigated by means of Magnetic Resonance Imaging (MRI). The aim of this study was to determine the local flow velocities at the inner face of the biofilm carrier. To get an insight into the mass transport processes, flow velocity maps of blank and with biofilm cultivated carriers were measured. A single carrier was placed in a tube in three different orientations and exposed to flow velocities of 0.21, 0.42, and 0.64 mm/s. The interplay of the biofilm morphology and the local flow pattern was then analyzed including the effect of the orientation of the carrier in relation to the upstream flow angle. Within this study, the biofilm carrier can be understood as an interconnected system of four sections in which the incoming fluid volume will be distributed depending on the biomass occupation and morphology. In sections with high biofilm occupation, the flow resistance is increased. Depending on the orientation of the carrier in the flow field, this effect leads to flow evasion through less covered sections showing higher flow velocities and consequently the risk of biofilm detachment. However, there was no clear correlation between biofilm coverage and flow ratio.

Low temperature partial nitritation/anammox in a moving bed biofilm reactor treating low strength wastewater.

Municipal wastewater collected in areas with moderate climate is subjected to a gradual temperature decrease from around 20 °C in summer to about 10 °C in winter. A lab-scale moving bed biofilm reactor (MBBR) with carrier material (K3 from AnoxKaldnes) was used to test the tolerance of the overall partial nitritation/anammox process to this temperature gradient. A synthetic influent, containing only ammonium and no organic carbon was used to minimize denitrification effects. After stable reactor operation at 20 °C, the temperature was slowly reduced by 2 °C per month and afterward held constant at 10 °C. Along the temperature decrease, the ammonium conversion dropped from an average of 40 gN m(-3) d(-1) (0.2 gN kgTSS h(-1)) at 20 °C to about 15 gN m(-3) d(-1) (0.07 gN kg TSS h(-1)) at 10 °C, while the effluent concentration was kept <8 mg NH4-N l(-1) during the whole operation. This also resulted in doubling of the hydraulic retention time over the temperature ramp. The MBBR with its biofilm on 10 mm thick carriers proved to sufficiently sustain enough biomass to allow anammox activity even at 10 °C. Even though there was a minor nitrite-build up when the temperature dropped below 12.5 °C, reactor performance recovered as the temperature decrease continued. Microbial community analysis by 16S rRNA amplicon analysis revealed a relatively stable community composition over the entire experimental period.