Integrated Fixed-film Activated Sludge Research Articles

Mechanisms of partial denitrification in an integrated fixed-film activated sludge (IFAS) system: From progressive startup to stabilization

The rapid startup and long-term stability of partial denitrification (PD) processes are essential prerequisites for successful PD/Anammox systems. Bioaggregates play a significant role in impacting mass transfer efficiency and shaping the microbial community structure within the PD process. Thus, the startup and stable operation of the PD process were investigated in an integrated fixed-film activated sludge (IFAS) by inoculating activated sludge and biofilm from anoxic tanks of a wastewater treatment plant. During the initial stage of startup, a sudden increase in NO3−-N concentration induced a rapid enzyme stress response, leading to a significant increase in total NaR activity (1.38 ± 0.0 μmol·(L·min)−1 to 25.94 ± 0.23 μmol·(L·min)−1), concomitant with the inhibition of total NiR activity (13.93 ± 0.48 μmol·(L·min)−1 to 4.25 ± 0.26 μmol·(L·min)−1), thereby inducing the rapid accumulation of NO2−-N (13.39 mg·L−1) within 6 days. During the later stage of startup, enzyme stress response was subsided but the partial denitrifiers were gradually enriched, consequently leading to continuous accumulation of NO2−-N. The results of ex-situ activity showed that flocs exhibit superior PD performance in contrast to biofilm throughout the entire operational period. Meanwhile, the total extracellular polymeric substances (EPS) content and community structure in both flocs and biofilm indicated that the migration of EPS-secreted partial denitrifiers (Thauera) from flocs to biofilm was obliged by substrate deficiency during the stable operating stage (days 35–100). Therefore, the startup of PD may be sequentially mediated by two distinct mechanisms, enzyme stress response and functional bacterial enrichment, with microbial community migration occurring during the stable phase.

Comprehensive comparison of integrated fixed-film activated sludge (IFAS) and AAO activated sludge methods: Influence of different operational parameters

Due to limited land availability in municipal wastewater treatment plants, integrated fixed-film activated sludge (IFAS) technology offers significant advantages in improving nitrogen removal performance and treatment capacity. In this study, two systems, IFAS and Anaerobic-Anoxic-Oxic Activated sludge process (AAO), were compared by adjusting parameters such as hydraulic retention time (HRT), nitrifying solution recycle ratio, sludge recycle ratio, and dissolved oxygen (DO). The objective was to investigate pollutant removal capacity and differences in microbial community composition between the two systems. The study showed that, at an HRT of 12 h, the IFAS system exhibited an average increase of 5.76%, 8.85%, and 12.79% in COD, NH4+-N, and TN removal efficiency respectively, compared to the AAO system at an HRT of 16 h. The TP concentration in the IFAS system reached 0.82 mg/L without the use of additives. The IFAS system demonstrated superior effluent results under lower operating conditions of HRT, nitrification solution recycle ratio, and DO. The 16S rDNA analysis revealed higher abundance of denitrification-related associated flora, including Proteobacteria, Bacteroidetes, and Planctomycetota, in the IFAS system compared to the AAO system. Similarities were observed between microorganisms attached to the media and activated sludge in the anaerobic, anoxic, and oxic tanks. q-PCR analysis indicated that the incorporation of filler material in the IFAS system resulted in similar abundance of nitrifying bacteria genes on the biofilm as in the oxic tank. Additionally, denitrifying genes showed higher levels due to aeration scouring and the presence of alternating aerobic-anaerobic environments on the biofilm surface, enhancing nitrogen removal efficiency.

Comparing methanol and glycerol as carbon sources for mainstream partial denitrification/anammox in an IFAS process.

This study explored the implementation of mainstream partial denitrification with anammox (PdNA) in the second anoxic zone of a wastewater treatment process in an integrated fixed film activated sludge (IFAS) configuration. A pilot study was conducted to compare the use of methanol and glycerol as external carbon sources for an IFAS PdNA startup, with a goal to optimize nitrogen removal while minimizing carbon usage. The study also investigated the establishment of anammox bacteria on virgin carriers in IFAS reactors without the use of seeding, and it is the first IFAS PdNA startup to use methanol as an external carbon source. The establishment of anammox bacteria was confirmed in both reactors 102 days after startup. Although the glycerol-fed reactor achieved a higher steady-state maximum ammonia removal rate because of anammox bacteria (1.6 ±0.3g/m2/day) in comparison with the methanol-fed reactor (1.2±0.2g/m2/day), both the glycerol- and methanol-fed reactors achieved similar average in situ ammonia removal rates of 0.39 ±0.2g/m2/day and 0.40 ±0.2g/m2/day, respectively. Additionally, when the upstream ammonia versus NOx (AvN) control system maintained an ideal ratio of 0.40-0.50 g/g, the methanol-fed reactor attained a lower average effluent TIN concentration (3.50 ±1.2mg/L) than the glycerol-fed reactor (4.43 ±1.6mg/L), which was prone to elevated nitrite concentrations in the effluent. Overall, this research highlights the potential for PdNA in IFAS configurations as an efficient and cost-saving method for wastewater treatment, with methanol as a viable carbon source for the establishment of anammox bacteria. PRACTITIONER POINTS: Methanol is an effective external carbon source for an anammox startup that avoids the need for costly alternative carbon sources. The methanol-fed reactor demonstrated higher TIN removal compared with the glycerol-fed reactor because of less overproduction of nitrite. Anammox bacteria was established in an IFAS reactor without seeding and used internally stored carbon to reduce external carbon addition. Controlling the influent ammonia versus NOx (AvN) ratio between 0.40 and 0.50 g/g allowed for low and stable TIN effluent conditions.

Performance and life cycle assessment of moving-bed biofilm and integrated fixed-film reactors to upgrade activated sludge systems

Upgrading the capacity of existing wastewater treatment plants (WWTPs) is hindered by the limited space for extension. This matter is particularly crucial in developing countries where dense urban areas experience unexpectedly growing populations. Herein, we study the capacity upgrading of a WWTP working by a conventional activated sludge (CAS) system from 180,000 m3/d to 240,000 and 300,000 m3/d. The WWTP case study focuses on a large plant in Mansoura, Egypt, requiring capacity upgrades to accommodate population growth. For this purpose, we used mathematical modeling to simulate current and suggested WWTP designs. Moving bed biofilm reactor (MBBR) and integrated fixed film-activated sludge (IFAS) systems were designed to increase the WWTP capacity to 240,000 and 300,000 m3/d by the addition of plastic biofilm carrier to 25% and 50% of the aeration basins, respectively. IFAS outperformed conventional and MBBR systems in terms of BOD, COD, TSS, NH4, TP, and PO4, in addition to increased capacity. We moreover used life cycle assessment (LCA) to comprehend the environmental favorability of upgrading scenarios. The most affected environmental categories are global warming and eutrophication due to the operation stage (especially electricity). Global warming potential for CAS, MBBR, and IFAS (300,000 m3/d) are 2.19, 1.69, and 1.73 kg CO2 eq respectively. Eutrophication potential for CAS, MBBR, and IFAS (300,000 m3/d) are 0.0025, 0.0022, and 0.0017 kg PO4- eq respectively. We also estimated the amortization and running costs for various upgrading scenarios, providing an integrated analysis for decision-making purposes.

Research progress of anaerobic ammonium oxidation (Anammox) process based on integrated fixed-film activated sludge (IFAS).

The integrated fixed-film activated sludge (IFAS) process is considered one of the cutting-edge solutions to the traditional wastewater treatment challenges, allowing suspended sludge and attached biofilm to grow in the same system. In addition, the coupling of IFAS with anaerobic ammonium oxidation (Anammox) can further improve the efficiency of biological denitrification. This paper summarises the research progress of IFAS coupled with the anammox process, including partial nitrification anammox, simultaneous partial nitrification anammox and denitrification, and partial denitrification anammox technologies, and describes the factors that limit the development of related processes. The effects of dissolved oxygen, influent carbon source, sludge retention time, temperature, microbial community, and nitrite-oxidising bacteria inhibition methods on the anammox of IFAS are presented. At the same time, this paper gives an outlook on future research focus and engineering practice direction of the process.

Review of emerging technologies for nutrient removal in wastewater treatment

The burgeoning global population and industrial activities have significantly increased the generation of wastewater laden with nutrients, posing severe environmental and public health concerns. Traditional wastewater treatment methods often fall short in effectively removing nutrients like nitrogen and phosphorus, leading to eutrophication of water bodies and endangering aquatic ecosystems. In response, emerging technologies for nutrient removal in wastewater treatment have gained traction in recent years, offering innovative and efficient solutions to mitigate nutrient pollution. This comprehensive review explores the latest advancements in nutrient removal technologies, encompassing biological, physical, and chemical processes. Biological treatment methods, including activated sludge, sequencing batch reactors (SBRs), and membrane bioreactors (MBRs), have been extensively studied and optimized for nutrient removal. Novel biofilm-based systems, such as moving bed biofilm reactors (MBBRs) and integrated fixed-film activated sludge (IFAS), have demonstrated enhanced nutrient removal capabilities and resilience to fluctuations in wastewater composition. Furthermore, the integration of advanced oxidation processes (AOPs) and membrane technologies has revolutionized nutrient removal from wastewater. AOPs, such as ozonation, ultraviolet (UV) irradiation, and photocatalysis, offer effective means to degrade recalcitrant organic pollutants and disrupt nutrient cycles. Membrane-based technologies, including reverse osmosis (RO), nanofiltration (NF), and forward osmosis (FO), enable selective nutrient removal and concentration, thereby producing high-quality effluent suitable for reuse or discharge into sensitive environments. Additionally, the review delves into emerging chemical treatment strategies, such as adsorption, precipitation, and ion exchange, for targeted removal of nutrients from wastewater streams. Advanced adsorbents and nanomaterials exhibit superior adsorption capacities and selectivity for nitrogen and phosphorus compounds, paving the way for cost-effective nutrient recovery and resource recycling. Moreover, the review highlights the importance of process optimization, system integration, and environmental sustainability in the development and deployment of emerging nutrient removal technologies. Life cycle assessments (LCAs) and techno-economic analyses provide valuable insights into the environmental footprint and economic viability of these innovative solutions, guiding decision-makers towards sustainable wastewater management practices. In conclusion, the synthesis of biological, physical, and chemical processes in emerging nutrient removal technologies holds great promise for addressing the challenges of nutrient pollution in wastewater treatment. Future research directions should focus on scalability, energy efficiency, and holistic approaches towards achieving water quality goals and fostering a circular economy.

Development and modeling of an integrated fixed-film activated sludge (IFAS) system for simultaneous nitrogen and carbon removal from an industrial estate wastewater

The potential of an integrated fixed film activated sludge (IFAS) bioreactor for developing simultaneous aerobic and anoxic micro-zones under continuous aeration regime to promote carbon and nitrogen removal from Faraman industrial estate wastewater was evaluated in the present research. The effects of three independent variables on carbon and nitrogen removal were assessed. Overall, the optimum condition with 94 %, 77 %, and 2 NTU of COD (chemical oxygen demand) removal, Total nitrogen (TN) removal, and effluent turbidity has been specified with hydraulic retention time (HRT) of 11 h, air flow rate (AFR) of 3.5 L/min, and filling ratio (FR) of 50 %. To assess the stability of treating processes in the system, the IFAS system was operated in this optimal condition. Moreover, the simulation of the bioreactor was accomplished via calibration and verification of GPS-X model. GPSX simulation results and experimental data were compared using an independent sample T-test, which the T-test result confirmed that there was no significant difference between them.

Performance evaluation of the integrated fixed-film activated sludge (IFAS) for industrial wastewater treatment containing ethylene glycol

Performance evaluation of the integrated fixed-film activated sludge (IFAS) for industrial wastewater treatment containing ethylene glycol

Deciphering the role of hydraulic retention time (HRT) in integrated fixed-film activated sludge (IFAS) system under aniline stress: Effects on microbial succession and communication

Integrated fixed-film activated sludge (IFAS) system offered an energy-efficient option for nitrogen removal from aniline wastewater. The regulation of hydraulic retention time (HRT) has been reported to affect the fate of functional microbial communities with different generation cycles, but the role of HRT in IFAS under aniline stress has not been sufficiently revealed. Herein, it was observed that shortening the HRT altered the mechanisms underlying the adaptation of a continuous flow micro-aerated IFAS system to aniline stress, thereby negligibly impacting the removal of aniline. The total nitrogen removal rate in Y2 (HRT = 0.5 day) was 40.28 % higher than Y1 (1 day), and 23.7 % higher than Y3 (0.25 day). This could be attributed to Y2 having an ample supply of electron donors for denitrification compared to Y1, as well as showing weaker stress to inhibit nitrification compared to Y3. Furthermore, with the progressive intensification of stress induced by HRT reduction, microorganisms gradually enhanced their adaptation to aniline. This led to an increased migration of microorganisms towards the inner layers of the biofilm, resulting in enhanced functional complementarity between the sludge and biofilm in Y2. Also, there was a distinct development of autotrophic denitrification, with arenimonas predominant in Y1, and Thauera predominant in Y2 and Y3. Moreover, Y2 possessed a high abundance of functional genes related to the production and reception of quorum sensing signal molecules, which may enhance the electron transport-dependent nitrogen metabolism reaction.

Stimulating extracellular polymeric substances production in integrated fixed-film activated sludge reactor for advanced nitrogen removal from mature landfill leachate via one-stage double anammox

Introducing carbon sources to achieve nitrogen removal from mature landfill leachate not only increases the costs and carbon emissions but also inhibits the activity of autotrophic bacteria. Thus, this study constructed a double anammox system that combines partial nitrification-anammox (PNA) and endogenous partial denitrification-anammox (EPDA) within an integrated fixed-film activated sludge (IFAS) reactor. In this system, PNA primarily contributes to nitrogen removal pathways, achieving a nitrite accumulation rate of 98.23%. The production of extracellular polymer substances (EPS) in the IFAS reactor is stimulated by introducing co-fermentation liquid. Through the utilization of EPS, the system effectively achieves EPDA with the nitrite transformation rate of 97.20%. Under the intermittent aeration operation strategy, EPDA combined with PNA and anammox in the oxic and anoxic stages enhanced the nitrogen removal efficiency of the system to 99.70 ± 0.12%. The functional genus Candidatus kuenenia became enriched in biofilm sludge, while Thauera and Nitrosomonas predominated in floc sludge.

Efficient nitrogen removal and substrate usage in integrated fixed-film activated sludge-anammox system under seasonal temperature variation

To elucidate how integrated fixed-film activated sludge (IFAS) system favors nitrogen removal performance under seasonal temperature variations, two push-flow reactors were operated with and without carriers under the same operating conditions. The results show that the IFAS system had significant advantages in shock response and low temperature adaptation, with a nitrogen removal rate of 0.37–0.53 kg-N(m3·d)-1 at the temperature of 8–12 °C. Anammox bacteria on carriers were almost unaffected by temperature variation, and its nitrogen removal contribution rate stabilized at 55 % in the IFAS system. The Haldane model reveals that the specific anammox activity in the IFAS system was 28 % to 49 % higher than that in the control system at 13 °C. Candidatus_Jettenia, with the highest abundance of 45 %, was the dominant species in the IFAS system and preferred to attach to the carriers. This study provides a feasible scheme for the application of anammox process.

Sequential adaptation strategies of SPDA systems to low temperature: EPS mediation and community structure evolution

Temperature stands as a vital factor in the practical application of simultaneous partial-denitrification/anammox (SPDA) systems in mainstream wastewater treatment, thus it is necessary to understand the adaptation strategy of SPDA systems to low temperature. To address this, the potential adaptation of different coupling forms to low temperatures was investigated in a moving bed biofilm reactor (MBBR) and an integrated fixed-film activated sludge (IFAS) reactor for 182 days. The results showed that IFAS reactor achieved higher total nitrogen removal rates (TNRRs) at 15 °C (89.67 ± 0.01%) than MBBR system (77.28 ± 0.02%). Nitrogen conversion in the typical cycles showed that PD process was more sensitive to low temperatures than anammox process in SPDA systems. Meanwhile, the significant increase of total extracellular polymeric substances (EPS) content in each biophase and the change of community structure indicated that EPS mediation and community structure evolution in different biophases are two adaptive strategies of SPDA system to temperature decreasing. Biofilm phase tends to produce more EPS to resist the temperature decrease than floc phase. The highly negative relationship between EPS content at 15 °C and the initial temperature of change in community structure (p = 0.036＜0.05, r = -0.964) suggested that microorganisms always tend to resist the adverse environment through EPS mediation first, and then adapt to the low-temperature environment through community structure evolution with the temperature further decreased. This study guided the selection of SPDA coupling system in practical engineering applications, and laid a foundation for efficient biological nitrogen removal at low temperatures.

Unveiling the roles of biofilm in reducing N2O emission in a nitrifying integrated fixed-film activated sludge (IFAS) system.

Biofilm process such as integrated fixed-film activated sludge (IFAS) system has been preliminarily found to produce less nitrous oxide (N2O) than suspended sludge system. However, the N2O emission behaviors and underlying N2O mitigation mechanism in such hybrid system remain unclear. This study therefore aims to fully unveil the roles of biofilm in reducing N2O emission in a nitrifying IFAS system with the aid of some advanced technologies such as N2O microsensor and site-preference analysis. It was found that ammonia oxidation occurred mostly in the sludge flocs (˃ 86%) and biofilm could reduce N2O emission by 43.77% in a typical operating cycle. Biofilm not only reduced nitrite accumulation in nitrification process, inhibiting N2O production via nitrifier denitrification pathway, but also served as a N2O sink, promoting the reduction of N2O via endogenous denitrification. As a result, N2O emissions from the IFAS system were 50%-83% lower than those from the solo sludge flocs. Further, more N2O emission was reduced in the presence of biofilm with decreasing the dissolved oxygen level in the range of 0.5-3.0 mg O2/L. Microbial community and key enzyme analyses revealed that biofilm had relatively high microbial diversity and unique enzyme composition, providing a reasonable explanation for the changed contributions of different N2O production pathways and reduced N2O emission.

Operational strategies enhancing sewage sludge minimization in a combined integrated fixed-film activated sludge – oxic settling anaerobic system

A lab-scale integrated fixed-film activated sludge (IFAS) reactor was mplemented with the oxic-settling anaerobic (OSA) cycle for reducing sewage sludge production through the addition of an anoxic/anaerobic sludge holding tank (SHT) along the sludge recycle line. The IFAS-OSA system was operated under the different hydraulic retention time (HRT) in the SHT (HRTSHT) of 12 h and 6 h, at an oxidation-reduction potential (ORP) < −91 mV and solid retention time (SRT) between 39 and 126 d. Furthermore, the effect of temperature increase in the SHT (TSHT) from ambient (19.8–25.6 °C) to mesophilic (35 °C) conditions was investigated. The system performances were monitored in terms of sludge minimization and dewaterability efficiencies as well as carbon and nutrients reduction. The observed sludge yield (Yobs) for the IFAS system was 0.37(±0.06) mg VSS/mg COD. After OSA implementation Yobs decreased by 32% and 46–65% at HRTSHT of 12 h and 6 h, respectively, indicating that prolonged exposure to anoxic/anaerobic conditions was not beneficial for sludge reduction. The lowest Yobs of 0.09(±0.05) mg VSS/mg COD (76% lower than that in the IFAS system) was obtained at an HRTSHT of 6 h and when TSHT was set at 35 °C. OSA implementation did not affect COD and NH4+ oxidation of the IFAS system (90–96% and 99%, respectively) and improved total nitrogen (TN) reduction (31–53%) due to improved denitrification in the SHT. On the contrary, sludge dewaterability worsened following OSA implementation, which was linked to the increased levels of exopolymeric substances in the suspended biomass.

Effect of dissolved oxygen concentration on performance and mechanism of simultaneous nitrification and denitrification in integrated fixed-film activated sludge sequencing batch reactors

Integrated fixed-film activated sludge (IFAS) is a superior system for achieving simultaneous nitrification and denitrification (SND), however, the impact of dissolved oxygen (DO) has not been fully elucidated. Therefore, this study investigated the effect of DO concentration on performance and mechanism of SND in IFAS system. Results showed that IFAS outperformed control systems and achieved optimal SND performance at a DO concentration of 0.5 mg/L, with an SND efficiency of 88.51% and total nitrogen removal efficiency of 82.78%. Typical cycles analysis demonstrated limited-DO promoted SND performance. Further analysis implied biofilms exhibited high biomass and denitrification activity with decreasing DO. Microbial community analysis revealed low DO concentrations were responsible for abundant functional groups and genes associated with SND and promoted unconventional nitrogen removal pathways. Moreover, co-occurrence network analysis elucidated microbial interactions, responses to DO, and keystone genera. This study helps understanding the roles of DO for enhanced SND in IFAS.

Effect of Operating Parameters on the Performance of Integrated Fixed-Film Activated Sludge for Wastewater Treatment.

Integrated fixed-film activated sludge (IFAS) is a hybrid wastewater treatment process that combines suspended and attached growth. The current review provides an overview of the effect of operating parameters on the performance of IFAS and their implications for wastewater treatment. The operating parameters examined include hydraulic retention time (HRT), solids retention time (SRT), dissolved oxygen (DO) levels, temperature, nutrient loading rates, and aeration. Proper control and optimization of these parameters significantly enhance the treatment efficiency and pollutant removal. Longer HRT and appropriate SRT contribute to improved organic matter and nutrient removal. DO levels promote the growth of aerobic microorganisms, leading to enhanced organic matter degradation. Temperature influences microbial activity and enzymatic reactions, impacting treatment efficiency. Nutrient loading rates must be carefully managed to avoid system overload or inhibition. Effective aeration ensures uniform distribution of wastewater and biofilm carriers, optimizing contact between microorganisms and pollutants. IFAS has been used in water reuse applications, providing a sustainable and reliable water source for non-potable uses. Overall, IFAS has proven to be an effective and efficient treatment process that can provide high-quality effluent suitable for discharge or reuse. Understanding the effects of these operating parameters helps to optimize the design and operation for efficient wastewater treatment. Further research is needed to explore the interactions between different parameters, evaluate their impact under varying wastewater characteristics, and develop advanced control strategies for improved performance and sustainability.

Partial nitritation-anammox for treatment of saline wastewater: Hydrazine-assisted salinity adaptation and nitrate control

In biological wastewater treatment, salinity can significantly restrict bacterial activity, and a long adaptation period to the saline condition is needed. This study investigated the feasibility of using partial nitritation-anammox (PNA) in an integrated fixed-film activated sludge (IFAS) reactor for enhanced biological nitrogen removal from saline municipal wastewater. The experimental results show that the salinity increase in the influent seriously affected the activity of anammox bacteria and disrupted the PNA process for autotrophic nitrogen removal when seawater was added to a 20% proportion with a resultant salinity of around 6.5‰. However, the addition of N2H4 (dosage: 5 mg/L for 3 weeks), an exogenous enhancer of both ammonia-oxidizing bacteria and anammox bacteria, effectively facilitated the rapid recovery and adaptation of PNA to salinity stress with a negligible additional cost for long-term operation. The PNA-IFAS reactor achieved a nitrogen removal capacity of 83.4 ± 5.9 mg N/(L‧d) in treating the saline municipal wastewater containing 40% seawater and an ammonia-N concentration of around 60 mg N/L. Salinity in the influent advantageously suppressed the activity and growth of the co-existing nitrite-oxidizing bacteria (NOB). The relative abundances of NOB in the suspended sludge (0.15%) and biofilms (0.33%) were kept at low levels, maintaining a low nitrate yield of 6.6 ± 1.5% in the bioreactor. The result suggests that salinity in the influent may offer potential benefits for reducing the risk of nitrate build-up and increasing the robustness of the PNA process for sustainable wastewater treatment.

Linking morphological features to anammox communities in a partial nitritation and anammox (PN/A) biofilm reactor

Partial nitritation/anammox (PN/A) has been recognized as a cost-efficient process for wastewater nitrogen removal. The addition of carriers could help achieve biomass retention and enhance the treatment efficiency by forming the dense biofilm. However, accurately determining the abundance of anammox bacteria (AnAOB) to evaluate the biofilm development still remains challenging in practice without access to specialized facilities and experimental skills. In this study, we explored the feasibility of utilizing the morphological features of anammox biofilm as an indication of the biofilm development progression, and its correlation with microbial communities was also revealed. The time-series biofilms from an integrated fixed-film activated sludge (IFAS) system with stable PN/A performance were sampled representing the different biofilm development stages. The biofilm morphological features including color and texture were respectively quantified by red (R) coordinate and Local binary pattern (LBP) descriptor via image processing. Hierarchy clustering analysis proved that the extracted morphological descriptors could well distinguish the different stages (colonization, succession, and maturation) of biofilm development. The microbial community dynamics of time-series anammox biofilms were investigated using the amplicon sequence variant (ASV) analysis. Candidatus Brocadia, as the typical AnAOB, dominated in the whole communities of 16.3%–20.0%, moreover, the biofilm development was found to be driven by distinct Brocadia species. Linear regression evidenced that the Brocadia abundance could be directly correlated to the value of R and LBP, and the total variation of microbial communities could be significantly explained by the morphological features via redundancy analysis. This study demonstrates a new way to monitor the biofilm development by extracting the visible features of anammox aggregates, which can help facilitate the automated control of anammox-based bioprocess.

Integrated fixed-film activated sludge systems in continuous-flow and batch mode acclimated from low to high aniline concentrations: Performance, mechanism and metabolic pathways

Integrated fixed-film activated sludge (IFAS) system has considerable advantages in treating aniline wastewater economically and efficiently. However, the response mechanism of IFAS to aniline needs further study. Herein, IFAS in continuous-flow (CF-IFAS) and batch mode (B-IFAS) were set up to investigate it. The removal efficiency of aniline exceeded 99% under different stress intensities. At low stress intensity (aniline ≈ 200 mg/L), the total nitrogen removal efficiency of B-IFAS was approximately 37.76% higher than CF-IFAS. When the stress intensity increased (aniline ≥ 400 mg/L), both were over 82%. CF-IFAS was restrained by denitrification while nitrification in B-IFAS. The legacy effect of perturbation of B-IFAS made microflora quickly reach new stability. The closer interspecific relationship in B-IFAS and more key species: Leucobacter, Rhodococcus, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Ellin6067 and norank_f_NS9_marine_group. Metabolic and Cell growth and death were the most abundant metabolic pathways, resulting both systems the excellent pollutant removal and stability under high stress intensity.

Co-Occurrence and Cooperation between Comammox and Anammox Bacteria in a Full-Scale Attached Growth Municipal Wastewater Treatment Process.

Cooperation between comammox and anammox bacteria for nitrogen removal has been recently reported in laboratory-scale systems, including synthetic community constructs; however, there are no reports of full-scale municipal wastewater treatment systems with such cooperation. Here, we report intrinsic and extant kinetics as well as genome-resolved community characterization of a full-scale integrated fixed film activated sludge (IFAS) system where comammox and anammox bacteria co-occur and appear to drive nitrogen loss. Intrinsic batch kinetic assays indicated that majority of the aerobic ammonia oxidation was driven by comammox bacteria (1.75 ± 0.08 mg-N/g TS-h) in the attached growth phase, with minimal contribution by ammonia-oxidizing bacteria. Interestingly, a portion of total inorganic nitrogen (∼8%) was consistently lost during these aerobic assays. Aerobic nitrite oxidation assays eliminated the possibility of denitrification as a cause of nitrogen loss, while anaerobic ammonia oxidation assays resulted in rates consistent with anammox stoichiometry. Full-scale experiments at different dissolved oxygen (DO = 2 - 6 mg/L) setpoints indicated persistent nitrogen loss that was partly sensitive to DO concentrations. Genome-resolved metagenomics confirmed the high abundance (relative abundance 6.53 ± 0.34%) of two Brocadia-like anammox populations, while comammox bacteria within the Ca. Nitrospira nitrosa cluster were lower in abundance (0.37 ± 0.03%) and Nitrosomonas-like ammonia oxidizers were even lower (0.12 ± 0.02%). Collectively, our study reports for the first time the co-occurrence and cooperation of comammox and anammox bacteria in a full-scale municipal wastewater treatment system.