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During long-term extra-terrestrial missions, food is limited and waste is generated. By recycling valuable nutrients from this waste via regenerative life support systems, food can be produced in space. Astronauts' urine can, for instance, be nitrified by micro-organisms into a liquid nitrate fertilizer for plant growth in space. Due to stringent conditions in space, microbial communities need to be be defined (gnotobiotic); therefore, synthetic rather than mixed microbial communities are preferred. For urine nitrification, synthetic communities face challenges, such as from salinity, ureolysis, and organics. In this study, a synthetic microbial community containing an AOB (Nitrosomonas europaea), NOB (Nitrobacter winogradskyi), and three ureolytic heterotrophs (Pseudomonas fluorescens, Acidovorax delafieldii, and Delftia acidovorans) was compiled and evaluated for these challenges. In reactor 1, salt adaptation of the ammonium-fed AOB and NOB co-culture was possible up to 45mScm -1 , which resembled undiluted nitrified urine, while maintaining a 44±10mgNH 4 + -NL -1 d -1 removal rate. In reactor 2, the nitrifiers and ureolytic heterotrophs were fed with urine and achieved a 15±6mg NO 3 - -NL -1 d -1 production rate for 1% and 10% synthetic and fresh real urine, respectively. Batch activity tests with this community using fresh real urine even reached 29±3mgNL -1 d -1 . Organics removal in the reactor (69±15%) should be optimized to generate a nitrate fertilizer for future space applications., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

Human urine accounts for only a fraction of the sewage volume, but it contains the majority of valuable nutrient load in wastewater. In this study, synthetic urine was nitrified in a closed photo-bioreactor through photosynthetic oxygenation by means of a consortium of microalgae and nitrifying bacteria. In situ production of oxygen by photosynthetic organisms has the potential to reduce the energy costs linked to conventional aeration. This energy-efficient strategy results in stable urine for further nutrient recovery, while part of the nutrients are biologically recovered in the form of valuable biomass. In this study, urine was nitrified for the first time without conventional aeration at a maximum photosynthetic oxygenation rate of 160 mg O 2 gVSS -1 d -1 (VSS: volatile suspended solids). A maximum volumetric nitrification rate of 67 mg N L -1 d -1 was achieved on 12% diluted synthetic urine. Chemical oxygen demand (COD) removal efficiencies were situated between 44% and 83% at a removal rate of 24 mg COD gVSS -1 d -1 . After 180 days, microscopic observations revealed that Scenedesmus sp. was the dominant microalga. Overall, photosynthetic oxygenation for urine nitrification is promising as a highly electricity efficient approach for further nutrient recovery.

Cost-efficient biological treatment of warm nitrogenous wastewaters requires the development of thermophilic nitrogen removal processes. Only one thermophilic nitrifying bioreactor was described so far, achieving 200 mg N L(-1) d(-1) after more than 300 days of enrichment from compost samples. From the practical point of view in which existing plants would be upgraded, however, a more time-efficient development strategy based on mesophilic nitrifying sludge is preferred. This study evaluated the adaptive capacities of mesophilic nitrifying sludge for two linear temperature increase patterns (non-oscillating vs. oscillating), two different slopes (0.25 vs. 0.08 °C d(-1)) and two different reactor types (floc vs. biofilm growth). The oscillating temperature pattern (0.25 °C d(-1)) and the moving bed biofilm reactor (0.08 °C d(-1)) could not reach nitrification at temperatures higher than 46 °C. However, nitrification rates up to 800 mg N L(-1) d(-1) and 150 mg N g(-1) volatile suspended solids d(-1) were achieved at a temperature as high as 49 °C by imposing the slowest linear temperature increase to floccular sludge. Microbial community analysis revealed that this successful transition was related with a shift in ammonium oxidizing archaea dominating ammonia oxidizing bacteria, while for nitrite oxidation Nitrospira spp. was constantly more abundant than Nitrobacter spp.. This observation was accompanied with an increase in observed sludge yield and a shift in maximal optimum temperature, determined with ex-situ temperature sensitivity measurements, predicting an upcoming reactor failure at higher temperature. Overall, this study achieved nitrification at 49 °C within 150 days by gradual adaptation of mesophilic sludge, and showed that ex-situ temperature sensitivity screening can be used to monitor and steer the transition process., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Nitrification is a well-studied and established process to treat ammonia in wastewater. Although thermophilic nitrification could avoid cooling costs for the treatment of warm wastewaters, applications above 40 °C remain a significant challenge. This study tested the effect of salinity on the thermotolerance of mesophilic nitrifying sludge (34 °C). In batch tests, 5 g NaCl L(-1) increased the activity of aerobic ammonia-oxidizing bacteria (AerAOB) by 20-21 % at 40 and 45 °C. For nitrite-oxidizing bacteria (NOB), the activity remained unaltered at 40 °C, yet decreased by 83 % at 45 °C. In a subsequent long-term continuous reactor test, temperature was increased from 34 to 40, 42.5, 45, 47.5 and 50 °C. The AerAOB activity showed 65 and 37 % higher immediate resilience in the salt reactor (7.5 g NaCl L(-1)) for the first two temperature transitions and lost activity from 45 °C onwards. NOB activity, in contrast to the batch tests, was 37 and 21 % more resilient in the salt reactor for the first two transitions, while no difference was observed for the third temperature transition. The control reactor lost NOB activity at 47.5 °C, while the salt reactor only lost activity at 50 °C. Overall, this study demonstrates salt amendment as a tool for a more efficient temperature transition for mesophilic sludge (34 °C) and eventually higher nitrification temperatures.

With oxygen supply playing a crucial role in an oxygen-limited autotrophic nitrification/denitrification (OLAND) rotating biological contactor (RBC), its controlling factors were investigated in this study. Disc rotation speeds (1.8 and 3.6rpm) showed no influence on the process performance of a lab-scale RBC, although abiotic experiments showed a significant effect on the oxygenation capacity. Estimations of the biological oxygen uptake rate revealed that 85-89% of the oxygen was absorbed by the microorganisms during the air exposure of the discs. Indeed, increasing the disc immersion (50 to 75-80%) could significantly suppress undesired nitratation, on the short and long term. The presented results demonstrated that nitratation could be controlled by the immersion level and revealed that oxygen control in an OLAND RBC should be predominantly based on the atmospheric exposure percentage of the discs., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

A nitrifying sequential batch reactor operated under 2-day cyclic aerobic and anoxic conditions was pulse dosed with incremental sulfide concentrations during anoxic conditions. The nitrite-oxidizing bacteria were found to be more sensitive to sulfide than the ammonia oxidizers. A maximum of nitrite-N to (nitrite-N + nitrate-N) accumulation ratio of 0.75 was obtained at an initial pulse sulfide-S concentration of 45 mg/L under pH control at 7.5 ± 0.2 and fully mixing conditions. Total ammonium nitrogen was removed almost 100% at a removal rate of 0.73 ± 0.05 g/L·day, achieved during the aerobic days of the cycles. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) analyses indicated the shift in the ammonia- and nitrite-oxidizing populations triggered by sulfide addition, partial nitrification, and subsequent recovery to complete nitrification. Interestingly, archaeal amoA genes were retrieved under the conditions of sulfide addition. These results indicate that the pulse sulfide application can be used as a tool to accumulate nitrite, which is of importance for the subsequent anaerobic ammonium oxidation (anammox) process in the achievement of complete nitrogen removal. [ABSTRACT FROM AUTHOR]

Abstract: Strains of Nitrobacter mediate the second step in the nitrification process by oxidizing nitrite to nitrate. The phylogenetic diversity of the genus is currently not well investigated. In this study, a rep-PCR profile and the nearly complete 16S rRNA gene sequence of 30 strains, comprising a wide physiological as well as ecological diversity and encompassing representatives of the four species, were determined. The sequence diversity of the 16S rRNA gene between different species was low, indicating the need for additional phylogenetic markers. Therefore, primers were developed for amplifying the complete nxrX gene and a 380bp fragment of the nxrB1 gene, which are both genes involved in the nitrite oxidation process. These genes confirmed the division into phylogenetic groups revealed by the 16S rRNA gene but showed a better discriminatory power. They can be a valuable additional tool for phylogenetic analysis within the genus Nitrobacter and can assist in the identification of new Nitrobacter isolates. [Copyright &y& Elsevier]

Gene sequence analysis of nirS and nirK, both encoding nitrite reductases, was performed on cultivated denitrifiers to assess their incidence in different bacterial taxa and their taxonomical value. Almost half of the 227 investigated denitrifying strains did not render an nir amplicon with any of five previously described primers. NirK and nirS were found to be prevalent in Alphaproteobacteria and Betaproteobacteria, respectively, nirK was detected in the Firmicutes and Bacteroidetes and nirS and nirK with equal frequency in the Gammaproteobacteria. These observations deviated from the hitherto reported incidence of nir genes in bacterial taxa. NirS gene phylogeny was congruent with the 16S rRNA gene phylogeny on family or genus level, although some strains did group within clusters of other bacterial classes. Phylogenetic nirK gene sequence analysis was incongruent with the 16S rRNA gene phylogeny. NirK sequences were also found to be significantly more similar to nirK sequences from the same habitat than to nirK sequences retrieved from highly related taxa. This study supports the hypothesis that horizontal gene transfer events of denitrification genes have occurred and underlines that denitrification genes should not be linked with organism diversity of denitrifiers in cultivation-independent studies. [ABSTRACT FROM AUTHOR]

Thermophilic nitrification has been proven in lab-scale bioreactors at 50 °C. The challenge is now to develop a solution for thermophilic nitrogen removal, integrating nitrification with denitrification and aerobic carbon removal. This pioneering study aimed at a single-sludge nitrification/denitrification process at 50 °C, through exposing nitrification in a step by step approach to anoxia and/or organics. Firstly, recurrent anoxia was tolerated by a nitrifying community during long-term membrane bioreactor (MBR) operation (85 days), with high ammonium oxidation efficiencies (>98%). Secondly, five organic carbon sources did not affect thermophilic ammonium and nitrite oxidation rates in three-day aerobic batch flask incubations. Moving to long-term tests with sequencing batch reactors (SBR) and MBR (>250 days), good nitrification performance was obtained at increasing COD/N influent ratios (0, 0.5, 1, 2 and 3). Thirdly, combining nitrification, recurrent anoxia and presence of organic carbon resulted in a nitrogen removal efficiency of 92–100%, with a COD/N removed of 4.8 ± 0.6 and a nitrogen removal rate of 50 ± 14 mg N g−1 VSS d−1. Overall, this is the first proof of principle thermophilic nitrifiers can cope with redox fluctuations (aerobic/anoxic) and the aerobic or anoxic presence of organic carbon, can functionally co-exist with heterotrophs and that single-sludge nitrification/denitrification can be achieved. Image 1 • The concept of single-sludge nitrification/denitrification was demonstrated at 50 °C • Thermophilic nitrifiers can cope with redox fluctuations (aerobic/anoxic) • Thermophilic nitrifiers can functionally co-exist with heterotrophs • 92–100% nitrogen removal was achieved at COD/N removed 4.8 ± 0.6 [ABSTRACT FROM AUTHOR]

Abstract: Three identical sequential batch reactors (SBRs) were each inoculated with sludge from a full-scale wastewater treatment plant (WWTP) treating a waste stream of different origin, i.e. a hospital, a meat processing company, and a municipal WWTP. The SBRs were run in parallel for 84 consecutive days to investigate whether the reactors would become more phylogenetically similar or stay separated concerning their functionality and microbial communities. Overall, the nitrification functionality was high throughout the experiment, and the size and structure of the sludge flocs were very similar. The total bacterial and ammonia-oxidizing bacterial (AOB) communities were analyzed by PCR-DGGE. Cluster analysis demonstrated very distinct bacterial communities in the three SBRs, not showing any trend becoming more similar. The carrying capacity, dynamics and functional organization of the communities were assessed by DGGE analysis and based on these patterns the range-weighted richness, moving window analysis, and constructing Pareto–Lorenz evenness distribution curves were calculated. Between the SBRs, highly comparable internal structure and dynamics of the AOB communities were observed, although they had only one AOB DGGE band in common. These observations indicate that community characteristics such as the extent of biodiversity and dynamics are more important indicators of good microbial functionality than the presence of certain specific species. [Copyright &y& Elsevier]

Abstract: The autotrophic two-species biofilm from the packed bed reactor of a life-support system, containing Nitrosomonas europaea ATCC 19718 and Nitrobacter winogradskyi ATCC 25391, was analysed after 4.8 years of continuous operation performing complete nitrification. Real-time quantitative polymerase chain reaction (Q-PCR) was used to quantify N. europaea and N. winogradskyi along the vertical axis of the reactor, revealing a spatial segregation of N. europaea and N. winogradskyi. The main parameters influencing the spatial segregation of both nitrifiers along the bed were assessed through a multi-species one-dimensional biofilm model generated with AQUASIM software. The factor that contributed the most to this distribution profile was a small deviation from the flow pattern of a perfectly mixed tank towards plug-flow. The results indicate that the model can estimate the impact of specific biofilm parameters and predict the nitrification efficiency and population dynamics of a multispecies biofilm. [Copyright &y& Elsevier]

Urine is a major waste product of human metabolism and contains essential macro- and micronutrients to produce edible microorganisms and crops. Its biological conversion into a stable form can be obtained through urea hydrolysis, subsequent nitrification, and organics removal, to recover a nitrate-enriched stream, free of oxygen demand. In this study, the utilization of a microbial community for urine nitrification was optimized with the focus for space application. To assess the role of selected parameters that can impact ureolysis in urine, the activity of six ureolytic heterotrophs (Acidovorax delafieldii, Comamonas testosteroni, Cupriavidus necator, Delftia acidovorans, Pseudomonas fluorescens, and Vibrio campbellii) was tested at different salinities, urea, and amino acid concentrations. The interaction of the ureolytic heterotrophs with a nitrifying consortium (Nitrosomonas europaea ATCC 19718 and Nitrobacter winogradskyi ATCC 25931) was also tested. Lastly, microgravity was simulated in a clinostat utilizing hardware for in-flight experiments with active microbial cultures. The results indicate salt inhibition of the ureolysis at 30 mS cm−1, while amino acid nitrogen inhibits ureolysis in a strain-dependent manner. The combination of the nitrifiers with C. necator and V. campbellii resulted in a complete halt of the urea hydrolysis process, while in the case of A. delafieldii incomplete nitrification was observed, and nitrite was not oxidized further to nitrate. Nitrate production was confirmed in all the other communities; however, the other heterotrophic strains most likely induced oxygen competition in the test setup, and nitrite accumulation was observed. Samples exposed to low-shear modeled microgravity through clinorotation behaved similarly to the static controls. Overall, nitrate production from urea was successfully demonstrated with synthetic microbial communities under terrestrial and simulated space gravity conditions, corroborating the application of this process in space. [ABSTRACT FROM AUTHOR]

Nitrification and denitrification, the key biological processes for thermophilic nitrogen removal, have separately been established in bioreactors at 50 °C. A well-characterized set of kinetic parameters is essential to integrate these processes while safeguarding the autotrophs performing nitrification. Knowledge on thermophilic nitrifying kinetics is restricted to isolated or highly enriched batch cultures, which do not represent bioreactor conditions. This study characterized the stoichiometry and kinetics of two thermophilic (50 °C) nitrifying communities. The most abundant ammonia oxidizing archaea (AOA) were related to the Nitrososphaera genus, clustering relatively far from known species Nitrososphaera gargensis (95.5% 16S rRNA gene sequence identity). The most abundant nitrite oxidizing bacteria (NOB) were related to Nitrospira calida (97% 16S rRNA gene sequence identity). The nitrification biomass yield was 0.20–0.24 g VSS g−1 N, resulting mainly from a high AOA yield (0.16–0.20 g VSS g−1 N), which was reflected in a high AOA abundance in the community (57–76%) compared to NOB (5–11%). Batch-wise determination of decay rates (AOA: 0.23–0.29 d−1; NOB: 0.32–0.43 d−1) rendered an overestimation compared to in situ estimations of overall decay rate (0.026–0.078 d−1). Possibly, the inactivation rate rather than the actual decay rate was determined in batch experiments. Maximum growth rates of AOA and NOB were 0.12–0.15 d−1 and 0.13–0.33 d−1 respectively. NOB were susceptible to nitrite, opening up opportunities for shortcut nitrogen removal. However, NOB had a similar growth rate and oxygen affinity (0.15–0.55 mg O 2 L−1) as AOA and were resilient towards free ammonia (IC 50 > 16 mg NH 3 -N L−1). This might complicate NOB outselection using common practices to establish shortcut nitrogen removal (SRT control; aeration control; free ammonia shocks). Overall, the obtained insights can assist in integrating thermophilic conversions and facilitate single-sludge nitrification/denitrification. Image 1 • Thermophilic nitrification yield was high, resulting mainly from a high AOA yield. • AOA belonged to the Nitrososphaera genus, NOB to Nitrospira calida. • Batch-wise determined decay rates were overestimated compared to in situ estimation. • NOB kinetics and high FA tolerance might complicate thermophilic NOB outselection. • Higher maximum temperature for AOA activity retention (55 °C) vs NOB (50 °C). [ABSTRACT FROM AUTHOR]

Even though nitrification/denitrification is a robust technology to remove nitrogen from sewage, economic incentives drive its future replacement by shortcut nitrogen removal processes. The latter necessitates high potential activity ratios of ammonia oxidizing to nitrite oxidizing bacteria (rAOB/rNOB). The goal of this study was to identify which wastewater and process parameters can govern this in reality. Two sewage treatment plants (STP) were chosen based on their inverse rAOB/rNOB values (at 20 °C): 0.6 for Blue Plains (BP, Washington DC, US) and 1.6 for Nieuwveer (NV, Breda, NL). Disproportional and dissimilar relationships between AOB or NOB relative abundances and respective activities pointed towards differences in community and growth/activity limiting parameters. The AOB communities showed to be particularly different. Temperature had no discriminatory effect on the nitrifiers' activities, with similar Arrhenius temperature dependences (Θ AOB = 1.10, Θ NOB = 1.06–1.07). To uncouple the temperature effect from potential limitations like inorganic carbon, phosphorus and nitrogen, an add-on mechanistic methodology based on kinetic modelling was developed. Results suggest that BP's AOB activity was limited by the concentration of inorganic carbon (not by residual N and P), while NOB experienced less limitation from this. For NV, the sludge-specific nitrogen loading rate seemed to be the most prevalent factor limiting AOB and NOB activities. Altogether, this study shows that bottom-up mechanistic modelling can identify parameters that influence the nitrification performance. Increasing inorganic carbon in BP could invert its rAOB/rNOB value, facilitating its transition to shortcut nitrogen removal. [ABSTRACT FROM AUTHOR]

Urine contains the majority of nutrients in urban wastewaters and is an ideal nutrient recovery target. In this study, stabilization of real undiluted urine through nitrification and subsequent microalgae cultivation were explored as strategy for biological nutrient recovery. A nitrifying inoculum screening revealed a commercial aquaculture inoculum to have the highest halotolerance. This inoculum was compared with municipal activated sludge for the start-up of two nitrification membrane bioreactors. Complete nitrification of undiluted urine was achieved in both systems at a conductivity of 75 mS cm −1 and loading rate above 450 mg N L −1 d −1 . The halotolerant inoculum shortened the start-up time with 54%. Nitrite oxidizers showed faster salt adaptation and Nitrobacter spp. became the dominant nitrite oxidizers. Nitrified urine as growth medium for Arthrospira platensis demonstrated superior growth compared to untreated urine and resulted in a high protein content of 62%. This two-stage strategy is therefore a promising approach for biological nutrient recovery. [ABSTRACT FROM AUTHOR]

The development of thermophilic nitrogen removal strategies will facilitate sustainable biological treatment of warm nitrogenous wastewaters. Thermophilic denitrification was extensively compared to mesophilic denitrification for the first time in this study. Two sequential batch reactors (SBR) at 34 °C and 55 °C were inoculated with mesophilic activated sludge (26 °C), fed with synthetic influent in a first phase. Subsequently, the carbon source was switched from acetate to molasses, whereas in a third phase, the nitrate source was fertilizer industry wastewater. The denitrifying sludge maintained its activity at 55 °C, resulting in an immediate process start-up, obtaining nitrogen removal rates higher than 500 mg N g-1 VSS d-1 in less than one week. Although the mesophilic SBR showed twice as high specific nitrogen removal rates, the maximum thermophilic denitrifying activity in this study was nearly 10 times higher than the activities reported thus far. The thermophilic SBR moreover had a 73% lower sludge volume index, a 45% lower sludge production and a higher resilience towards a change in carbon source compared with the mesophilic SBR. The higher resilience was potentially related to a higher microbial diversity and evenness of the thermophilic community at the end of the synthetic feeding period. The thermophilic microbial community showed a higher similarity over the different feeding periods implying a more stable community. Overall, this study showed the capability of mesophilic denitrifiers to maintain their activity after a large temperature increase. Existing mesophilic process systems with cooling for the treatment of warm wastewaters could thus efficiently be converted to thermophilic systems with low sludge production and good settling properties. [ABSTRACT FROM AUTHOR]

Ammonia gas is conventionally treated in nitrifying biofilters; however, addition of organic carbon to perform post-denitrification is required to obtain total nitrogen removal. Oxygen-limited autotrophic nitrification/denitrification (OLAND), applied in full-scale for wastewater treatment, can offer a cost-effective alternative for gas treatment. In this study, the OLAND application thus was broadened toward ammonia loaded gaseous streams. A down flow, oxygen-saturated biofilter (height of 1.5 m; diameter of 0.11 m) was fed with an ammonia gas stream (248 ± 10 ppmv) at a loading rate of 0.86 ± 0.04 kg N m-3 biofilter d-1 and an empty bed residence time of 14 s. After 45 days of operation a stable nitrogen removal rate of 0.67 ± 0.06 kg N m-3 biofilter d-1, an ammonia removal efficiency of 99%, a removal of 75-80% of the total nitrogen, and negligible NO/N2O productions were obtained at water flow rates of 1.3 ± 0.4 m3 m-2 biofilter section d-1. Profile measurements revealed that 91% of the total nitrogen activity was taking place in the top 36% of the filter. This study demonstrated for the first time highly effective and sustainable autotrophic ammonia removal in a gas biofilter and therefore shows the appealing potential of the OLAND process to treat ammonia containing gaseous streams. [ABSTRACT FROM AUTHOR]

This study assessed the technical feasibility to treat digested black water from vacuum toilets (> 1000 mg NH4+-NL-1) in a lab-scale oxygen-limited autotrophic nitrification/denitrification (OLAND) rotating biological contactor. After an adaptation period of 2.5 months, a stable nitrogen removal rate of ca. 700 mg NL-1d-1 was reached over the subsequent 5 months. Suppression at the nitrite oxidizing bacteria at free ammonia levels above 3 mg NL-1 resulted in a nitrogen removal efficiency of 76%. The favorable ratios of both organic and inorganic carbon to nitrogen guaranteed endured anammox activity and sufficient buffering capacity, respectively. Quantitative FISH showed that aerobic and anoxic ammonium-oxidizing bacteria (AerAOB and AnAOB) made up 43 and 8% of the biofilm, respectively. Since a part of the AerAOB was probably present in anoxic biofilm zones, their specific ammonium conversion was very low, in contrast to the high specific AnAOB activity. DGGE analysis showed that the dominant AerAOB and AnAOB species were resistant to the transition from synthetic medium to digested black water. This study demonstrates high-rate nitrogen removal from digested black water by one-stage partial nitritation and anammox, which will allow a significant decrease in operational costs compared to conventional nitrification/denitrification. [ABSTRACT FROM AUTHOR]

Flow-through reactors with manganese oxides were examined for their capacity to remove 17α-ethinylestradiol (EE2) at μg L−1 and ng L−1 range from synthetic wastewater treatment plant (WWTP) effluent. The mineral MnO2 reactors removed 93% at a volumetric loading rate (B V) of 5 μg EE2 L−1 d−1 and from a B V of 40 μg EE2 L−1 d−1 on, these reactors showed 75% EE2 removal. With the biologically produced manganese oxides, only 57% EE2 was removed at 40 μg EE2 L−1 d−1. EE2 removal in the ng L−1 range was 84%. The ammonium present in the influent (10 mg N L−1) was nitrified and ammonia-oxidizing bacteria (AOB) were found to be of prime importance for the degradation of EE2. Remarkably, EE2 removal by AOB continued for a period of 4 months after depleting NH4 + in the influent. EE2 removal by manganese-oxidizing bacteria was inhibited by NH4 +. These results indicate that the metabolic properties of nitrifiers can be employed to polish water containing EE2 based estrogenic activity. [Copyright &y& Elsevier]

A sequential batch reactor (SBR) and a membrane bioreactor (MBR) were inoculated with the same sludge from a municipal wastewater treatment plant, supplemented with ammonium, and operated in parallel for 84 days. It was investigated whether the functional stability of the nitrification process corresponded with a static ammonia-oxidizing bacterial (AOB) community. The SBR provided complete nitrification during nearly the whole experimental run, whereas the MBR showed a buildup of 0 to 2 mg nitrite-N liter-1 from day 45 until day 84. Based on the denaturing gradient gel electrophoresis profiles, two novel approaches were introduced to characterize and quantify the community dynamics and interspecies abundance ratios: (i) the rate of change [Δt(week)] parameter and (ii) the Pareto-Lorenz curve distribution pattern. During the whole sampling period, it was observed that neither of the reactor types maintained a static microbial community and that the SBR evolved more gradually than the MBR, particularly with respect to AOB (i.e., average weekly community changes of 12.6% ± 5.2% for the SBR and 24.6% ± 143% for the MBR). Based on the Pareto-Lorenz curves, it was observed that only a small group of AOB species played a numerically dominant role in the nitritation of both reactors, and this was true especially for the MBR. The remaining less dominant species were speculated to constitute a reserve of AOB which can proliferate to replace the dominant species. The value of these parameters in terms of tools to assist the operation of activated-sludge systems is discussed. [ABSTRACT FROM AUTHOR]

• Integrated fixed-film activated sludge tested for shortcut nitrogen removal. • Combining strategies selectively suppressed activity nitrite oxidizing bacteria. • Flocs acted as nitrite source, while carriers were nitrite sinks. • Optimal nitrite source: low, but sufficient aerobic floc retention time (±7 days) • Optimal sink: dissolved oxygen < 0.47 mg O 2 L−1 and N loading rate > 150 mg N L−1 d−1. Implementation of mainstream partial nitritation/anammox (PN/A) can lead to more sustainable and cost-effective sewage treatment. For mainstream PN/A reactor, an integrated fixed-film activated sludge (IFAS) was operated (26 °C). The effects of floccular aerobic sludge retention time (AerSRT floc), a novel aeration strategy, and N-loading rate were tested to optimize the operational strategy. The best performance was observed with a low, but sufficient AerSRT floc (~7d) and continuous aeration with two alternating dissolved oxygen setpoints: 10 min at 0.07–0.13 mg O 2 L−1 and 5 min at 0.27–0.43 mg O 2 L−1. Nitrogen removal rates were 122 ± 23 mg N L−1 d−1, and removal efficiencies 73 ± 13%. These conditions enabled flocs to act as nitrite sources while the carriers were nitrite sinks, with low abundance of nitrite oxidizing bacteria. The operational strategies in the source-sink framework can serve as a guideline for successful operation of mainstream PN/A reactors. [ABSTRACT FROM AUTHOR]

Anammox, the oxidation of ammonium with nitrite, is a key microbial process in the nitrogen cycle. Under mesophilic conditions (below 40 °C), it is widely implemented to remove nitrogen from wastewaters lacking organic carbon. Despite evidence of the presence of anammox bacteria in high-temperature environments, reports on the cultivation of thermophilic anammox bacteria are limited to a short-term experiment of 2 weeks. This study showcases the adaptation of a mesophilic inoculum to thermophilic conditions, and its characterization. First, an attached growth technology was chosen to obtain the process. In an anoxic fixed-bed biofilm bioreactor (FBBR), a slow linear temperature increase from 38 to over 48 °C (0.05–0.07 °C d−1) was imposed to the community over 220 days, after which the reactor was operated at 48 °C for over 200 days. Maximum total nitrogen removal rates reached up to 0.62 g N L−1 d−1. Given this promising performance, a suspended growth system was tested. The obtained enrichment culture served as inoculum for membrane bioreactors (MBR) operated at 50 °C, reaching a maximum total nitrogen removal rate of 1.7 g N L−1 d−1 after 35 days. The biomass in the MBR had a maximum specific anammox activity of 1.1 ± 0.1 g NH 4 +-N g−1 VSS d−1, and the growth rate was estimated at 0.075–0.19 d−1. The thermophilic cultures displayed nitrogen stoichiometry ratios typical for mesophilic anammox: 0.93–1.42 g NO 2 --N removed g−1 NH 4 +-N removed and 0.16–0.35 g NO 3 --N produced g−1 NH 4 +-N removed. Amplicon and Sanger sequencing of the 16S rRNA genes revealed a disappearance of the original "Ca. Brocadia" and "Ca. Jettenia" taxa, yielding Planctomycetes members with only 94–95% similarity to "Ca. Brocadia anammoxidans" and "Ca. B. caroliniensis", accounting for 45% of the bacterial FBBR community. The long-term operation of thermophilic anammox reactors and snapshot views on the nitrogen stoichiometry, kinetics and microbial community open up the development path of thermophilic partial nitritation/anammox. A first economic assessment highlighted that treatment of sludge reject water from thermophilic anaerobic digestion of sewage sludge may become attractive. Image 1 • A slow temperature increase on a mesophilic inoculum enabled thermophilic anammox. • A potentially novel species was highly abundant in the adapted community. • Stoichiometry and rescaled kinetics are low compared to mesophilic anammox cultures. • An attached and a suspended growth reactor were feasible. • Thermophilic partial nitritation/anammox may become economically cost-effective. [ABSTRACT FROM AUTHOR]