Nitrite-oxidizing bacteria (NOB) seriously threaten the partial nitritation and Anammox (PN/A) process, hindering its mainstream application. Herein, a one-stage PN/A reactor was continuously operated for 245 days under nitrogen loading rate lifted from 0.4 g N/L/d to 0.6 g N/L/d and 0.8 g N/L/d with the nitrogen removal efficiency of 71 %, 64 %, and 41 %, respectively. Furthermore, the NOB species over time was identified as Nitrospira_sp._OLB3, exhibiting an increase of the relative abundance from 0.9 % to 4.3 %. The hydroxyapatite (HAP) granules gradually lost their microbiological function of Anammox bacteria then aged, leading to NOB dominance. Therefore, one "pulse therapy" was introduced and combined with "continuous enhancement" of Anammox sludge supported by sidestream to competitively limit the NOB dynamics. The treatment's effect persisted for around two months. The strategy that returning at least 50 % of the impaired HAP granular sludge to the sidestream for recultivation could fulfill the bottlenecks of mainstream PN/A.

Anammox-based processes are attractive for biological nitrogen removal, and the combination of anammox and hydroxyapatite (HAP) is promising for the simultaneous removal of nitrogen and phosphorus from wastewater. However, the kinetics of one-stage partial nitritation/anammox (PNA) in which ammonia-oxidizing bacteria (AOB) and anammox bacteria (AnAOB) exist in a reactor are poorly understood. Moreover, inorganic elements are required to promote microbial cell synthesis and growth; therefore, monitoring of elements to prevent the limitation and inhibition of the process is critical. The minimum amounts of inorganic elements required for a one-stage PNA process and the elemental flow remain unknown. Therefore, in this study, kinetics, stoichiometry, and element flow in the long-term, high-rate, continuous, one-stage HAP-PNA process with microaerobic granular sludge at 25 °C were determined using process modeling, parameter estimation, and mass balance. The biomass elemental composition was determined to be CH2.2O0.89N0.18S0.0091, and the biomass yield (Yobs) was calculated to be 0.0805 g/g NH4+-N. Therefore, a stoichiometric reaction equation for the one-stage HAP-PNA system was also proposed. The maximum specific growth rate (μm) of AnAOB and AOB were 0.0360 and 0.0982 d-1 with doubling times of 19 and 7.1 d, respectively. Finally, the elemental requirements for stable and high-rate performance were determined using element flow analysis. These findings are essential for developing the anammox-based process in a stable and resource-efficient manner and determining engineering applicability.

The improvement of organic sludge destruction efficiency and methanogenic performance is a key concern during anaerobic digestion toward maximum energy recovery. In this study, a high-solid anaerobic membrane bioreactor (AnMBR) was operated continuously for the treatment of organic sludge from Japanese small-scale collective wastewater treatment facility (Johkasou), and digestion efficiency was enhanced by the optimizing solid retention time (SRT). Degradation efficiency of the substrate improved from 36 % to 52 % and the biogas yield was enhanced from 0.37 to 0.51 L/g-VSfed when the SRT was extended from 30 to 60 d. The net energy yield of AnMBR at SRT 60 days was 9.83 kJ/g-VSfed, and the corresponding energy sufficiency ratio was 181 %, indicating that SRT extension could enhance substrate destruction with significant energy recovery potential. However, a long SRT is characterized by high mixed liquor total solids (MLTS), small particle size, high extracellular polymeric substances content, and poor filterability, which exert detrimental effects on membrane operation. Membrane fouling was effectively controlled by regulating the flux at a sustainable rate. The low fouling region and transition region of operating flux were determined as 0.21-4.6 L/m2/h (LMH) and 1.5-5.7 LMH, respectively, when MLTS was 25-50 g/L, and the main contributors to membrane fouling were high protein fractions and small sludge flocs. The current study proposes a promising method to promote digestion efficiency and provided adequate guidance for membrane operation at super-high MLTS by presenting practical engineering applications of AnMBRs in solid waste treatment.

Retaining sufficient anammox bacteria (AnAOB) while keeping the anammox-based process stable is the focus of the study of anammox technology, especially in a one-stage partial nitritation/anammox (PNA) process. The use of hydroxyapatite (HAP) granules in an anammox-based process is innovative for its potential to improve the nitrogen removal rate and achieve simultaneous removal of phosphorus. In this study, the HAP-based granular sludge was employed using enhancement strategies for an excellent nitrogen removal performance in a one-stage PNA process. Compared to those of other granular sludge PNA systems, a remarkable sludge volume index of 7.8 mL/g and an extremely high mixed liquor volatile suspended solids of 15 g/L were achieved under a low hydraulic retention time of 2 h. Consequently, an unprecedented nitrogen removal rate as high as 4.8 kg N/m3/d at 25 °C was obtained under a nitrogen loading rate of 6 kg N/m3/d. After a long-term operation of 870 days, the enhancement strategies underlying the superior performance of the granular sludge were identified. These findings clearly demonstrate that the enhancement strategies are crucial for the superior operating performance of the PNA process, and they can promote the application of the anammox-based process.

High solid anaerobic membrane bioreactor (HSAnMBR) is widely applied in biomass treatment and energy regeneration, while membrane operation performance and membrane fouling control remain critical issues. In this study, a HSAnMBR was utilized for waste activated sludge (WAS) treatment at organic loading rates of 3.69-3.72 gCOD/L·d and biogas yield was ranged in 0.38-0.39 L/gVSfed with the COD conversion efficiency of 40 %. The membrane operated stably when the average flux was 9.6, 4.5 and 1.2 L/m2/h at mixed liquor total solid of 25, 30 and 40 g/L with a filtration: relaxation of 4:1, 1:1 and 1:2, respectively. The distinctive characteristics of membrane fouling at high solid condition were that the polysaccharides and proteins had high fouling propensity and were the main composition of the foulant layer. Furthermore, phosphorus and magnesium were the predominant causes of inorganic fouling. The minerals precipitated on the membrane and were embedded into membrane pores, contributing to cake layer formation and pore blocking. This research provided a comprehensive analysis of the membrane operation characterization and fouling mechanisms of HSAnMBR, which was expected to push forward HSAnMBR applications to WAS treatment.

The anaerobic ammonia oxidation (anammox) process is a promising biological nitrogen removal technology. However, owing to the sensitivity and slow cell growth of anammox bacteria, long startup time and initially low nitrogen removal rate (NRR) are still limiting factors of practical applications of anammox process. Moreover, nitrogen removal efficiency (NRE) is often lower than 88 %. This review summarizes the most common methods for improving NRR by increasing microorganism concentration, and modifying reactor configuration. Recent integrated anammox-based systems were evaluated, including hydroxyapatite (HAP)-enhanced one-stage partial nitritation/anammox (PNA) process for a high NRR of over 2 kg N/m3/d at 25 °C, partial denitrification/anammox (PDA) process, and simultaneous partial nitrification, anammox, and denitrification process for a high NRE of up to 100 %. After discussing the challenges for the application of these systems critically, a combined system of anaerobic digestion, HAP-enhanced one-stage PNA and PDA is proposed in order to achieve a high NRR, high NRE, and phosphorus removal simultaneously.

Nitrogen removal from wastewater by the partial nitritation/anammox (PNA) technology is promising from both economic and environmental perspectives. However, this technology has not been popularized in the mainstream because of low biomass retention and the growth of the nitrite oxidizing bacteria. In this study, a one-stage PNA process with hydroxyapatite (HAP)-enhanced granules was used to treat effluent from a mainstream anaerobic membrane bioreactor. The HAP-enhanced reactor allowed an enriched high biomass of 6.9 ± 0.2 g/L at a low hydraulic retention time of 2 h. A nitrogen removal efficiency of 80 ± 6.0 %, a nitrogen removal rate of 0.36 ± 0.05 kg/m3/d and a COD removal efficiency of 54 ± 15 % were achieved stably, leading to a low total nitrogen concentration of 8.5 ± 2.7 mg/L and a low COD concentration of 19.7 ± 5.9 mg/L in the effluent. Anammox bacteria of Candidatus Kuenenia stuttgartiensis and ammonium oxidizing bacteria of Nitrosomonas were found to be the two most predominant bacteria.

An integrated system combining in pipe thermal pretreatment with a high-solid anaerobic membrane bioreactor (AnMBR) was developed to promote the anaerobic digestion of waste activated sludge (WAS). Two different pretreatment methods investigated were the venturi nozzle treatment (VNT) and steam injector treatment (SIT), both at a low temperature of 70 °C. The biogas production after pretreatment was 23.5-30.5% higher than that of untreated WAS, and the VS based biogas yield was 0.46-0.47 L/g-VS when HRT was 15 days. The membrane operated smoothly when the average flux was 9.6 and 4.5 L/m2/h under an MLTS of 25 and 30 g/L, respectively. The calculations of the mass balance indicated that 44-45% COD was converted to methane with pretreatment and only 1% remained in the permeate. That is, high energy recovery and organic matter removal efficiency were achieved for the treatment of WAS using the high-solid AnMBR with in pipe thermal pretreatment.

The simultaneous partial nitritation, anammox, denitrification, and COD oxidation (SNADCO) method was successfully carried out in an air-lift moving bed biofilm reactor (AL-MBBR) with cylinders carriers for the treatment of digested fish processing wastewater (FPW). Synthetic wastewater was used as substrate at stage 1. It changed into the digested FPW with dilution variation in order to increase the nitrogen and COD loading rates. With influent concentration of NH4+-N of 909 ± 101 mg-N/L and COD of 731 ± 26 mg/L, the nitrogen removal efficiency was 86.8% (nitrogen loading rate of 1.21 g-TN/L/d) and the COD removal efficiency was 50.5% (COD loading rate at 0.98 g-COD/L/d). This study showed that the process has the advantages in treating the real high ammonia concentration of digested wastewater containing organic compounds. The nitritation and anammox route was predominant in nitrogen removal, while COD oxidation and microbe proliferation played the main role in COD removal.

The one-stage partial nitritation and anammox process with the hydroxyapatite (HAP)-based syntrophic granules was studied for the ammonium nitrogen removal from the effluents of a self-agitated anaerobic baffled reactor treating the fish processing wastewater. When the ammonium in the influent was 1140 mg N·L-1, a high nitrogen removal rate and nitrogen removal efficiency of 1.51 ± 0.10 kg N·m-3·d-1 and 88.2% were obtained, respectively. Anammox bacteria of Candidatus Kuenenia stuttgartiensis and ammonium oxidizing bacteria of Nitrosomonas were the two most predominant bacteria, while nitrite oxidizing bacteria activity was low and could be neglected during the treatment. The inorganic element properties of the sludge were analyzed by several methods to confirm the existence of HAP granules. Optical microscopic observation and scanning electron microscopy analysis revealed the structure of the granular sludge.This study supports the feasibility and potential of this process for high-efficiency nitrogen removal from fish processing wastewater.

In low-strength ammonium wastewater (LSAWW) treatment, the application of anammox-based process is still limited due to extreme instability and the poor nitrogen removal rate (NRR). In this work, granule sludge, comprised of functional microbes and hydroxyapatite (HAP), was inoculated and cultivated in a one-stage partial nitritation/anammox (PNA) reactor for LSAWW treatment. The results showed that at the hydraulic retention time (HRT) of about 1.0 h and the influent ammonium concentration of 63.0 mg/L, an average NRR of 1.28 kg/m3/d was achieved, which far exceeds that reported in similar studies. The main inorganic matter in sludge was identified as HAP through the X-ray diffractometer and Raman spectrum analysis. The tomographic images of wet granule created through computed tomography revealed that the interior density of the granules was uneven and many hollow structures existed in the granule interior. Combined with the Scanning Electron Microscope images of dry granules, it was found that the granules were comprised of hollow sub-granules. Since the biomass in the reactor increased with no obvious increase in the granule size, it was inferred that the hollow sub-granules had fragile connections with each other and that granules division occurred easily, resulting in the high dispersity of sludge. Florescence in situ hybridization results also showed that the ammonium-oxidizing bacteria and anammox bacteria were mainly distributed in the two sides of the sub-granule shells and the HAP in the middle. This kind of structure raised the density of granules and improved the settleability of sludge, which made it possible to achieve a high biomass in the reactor at a short HRT. Therefore, the sludge formed in the reactor was concentrated, highly dispersive and easily settleable. These factors appear to be crucial for achieving the desired nitrogen removal performance. This study marks a big leap in LSAWW treatment through the one-stage PNA process and has great potential in actual applications.

A 20 L hollow-fiber submerged anaerobic membrane bioreactor (SAnMBR) was used to treat real domestic wastewater at 25 °C with hydraulic retention times (HRTs) ranging from 4 to 12 h. The process performance was evaluated by organic removal efficiency, biogas production, sludge yield, and filtration behaviors during one-year's operation. For HRTs ranging between 6 and 12 h, the AnMBR showed good organic removal efficiency with chemical oxygen demand (COD) and biochemical oxygen demand (BOD) removal efficiencies of about 89% and 93%, respectively. The biogas yield was 0.26 L-gas/g-CODfed, with approximately 80% methane content, and the sludge yield was 0.07-0.11 g-VSS/g-CODrem. While at an HRT of 4 h, with the higher wastewater treatment capacity and organic loading rate (OLR), the biogas production was lower (0.17 L-gas/g-CODfed), and the sludge production was higher (0.22 g-VSS/g-CODrem). The organic removal performance (COD 84% and BOD 89%) at HRT of 4 h was acceptable due to the effective separation effect of the membrane filtration process. According to COD balance analysis, the low biogas yield and high sludge yield at HRT of 4 h were due to insufficient biodegradation under an OLR of 2.05 g-COD/L-reactor/d. Theoretical calculations based on Henry's law indicate that the ideal methane content in the biogas should be 82-85% when the operational temperature was 25 °C. To achieve a high flux and sustainable AnMBR operation, the impact of mixed liquor suspended solid (MLSS) and gas sparging velocity (GSV) on the filtration performance was analyzed. The critical flux increased with increase in the GSV from 24.2 to 174.3 m/h, but decreased with increase in the MLSS concentration from 8.2 to 20.2 g/L. Therefore, decreasing fouling rate to 0.8-1.2 kPa/d by efficiently controlling GSV and MLSS, sustainable operation could be achieved at a flux of 0.34 m/d.

Pore size is one of the most important properties in the successful operation of membrane-based bioprocesses for the treatment of municipal wastewater. The characteristics of two anaerobic membrane bioreactors (AnMBRs), one with a hollow fiber membrane of 0.4 μm pore size (AnMBR1), and the other with a membrane of 0.05 μm pore size (AnMBR2) were investigated for the treatment of real municipal wastewater at room temperature (25 °C) under varied hydraulic retention times (HRTs). Process performance was evaluated in terms of organic removal efficiency, biogas production and membrane filtration behaviours during a long-term continuous operation. Both AnMBRs showed good organic removal performance with COD and BOD removal efficiencies of around 89% and 93%, respectively. High energy recovery potential was achieved, with the biogas yield ranging between 0.20 and 0.26 L-gas/g-CODrem and a methane content of approximately 75%. The differences in the membrane filtration behaviours in the two AnMBRs included different permeate flux and total filtration resistance (Rt). In the AnMBR with a 0.4 μm pore size membrane, an average Rt of 1.08 × 10^12 m-1 was obtained even when the permeate flux was a high 0.274 m/day, while a higher average Rt of 1.51 × 10^12 m-1 was observed in the AnMBR with 0.05 μm pore size membrane even when the flux was a low 0.148 m/day. The off-line membrane cleaning strategy used for AnMBR1 indicated that the membrane restoration efficiency was 90.2%.

The freshwater anammox-based process would usually experience performance fluctuation and need adoption period when subjected to saline wastewater according to previous studies. In this study, the performance of nitrite-limited freshwater one-stage partial nitritation/anammox (PNA) process subjected to saline wastewater was investigated. The results showed that the nitrite-limited freshwater system could smoothly adapt to the salinity of 0.25%. The stable nitrogen removal could be achieved until the salinity of 2.5%, at which the desirable average NRR of 0.74 ± 0.1 kg/m3/d was achieved. The microbial community analysis showed that during the whole experiment, the main functional microbes were from genus Nitrosomonas and genus Kuenenia, which were through to be the crucial factors for achieving the excellent performance. This study indicates the nitrite-limited strategy is admissible for stabilizing the performance of freshwater one-stage PNA process subjected to saline wastewater in actual application.

Over the past forty years, anammox-based processes have been extensively researched and applied to some extent. However, some of the long-standing problems present serious impediments to wide application of these processes, and knowledge gap between lab-scale research and full-scale operations is still considerable. In recent years, anammox-based research has developed rapidly and some emerging concepts have been proposed. The focus of this review is on the critical problems facing actual application of anammox processes. The latest developments in anammox-based processes are summarized, and particular consideration is given to the following aspects: (1) the evolution of the chemical stoichiometry of anammox reaction; (2) the status of several main anammox-based processes; (3) the critical problems and countermeasures; (4) the emerging anammox-based processes; and (5) the suggested optimal process integrating partial nitritation, anammox, hydroxyapatite crystallization and denitratation for digestion effluent treatment towards more efficient nitrogen removal and phosphorus recovery in the future.

The stable operation of the partial nitritation and anammox (PN/A) process is a challenge in the treatment of low-strength ammonia wastewater like sewage mainstream. This study demonstrated the feasibility of achieving stable operation in the treatment of 50 mg/L ammonia wastewater with a micro granule-based PN/A reactor. The long-term operation results showed nitrogen removal efficiencies of 71.8 ± 9.9% were stably obtained under a relatively short hydraulic retention time (HRT) of 2 h. The analysis on the physicochemical properties of the granules indicated most of the granules were in a size in a range of 265-536 μm, and the elementary composition of the granules was determined to be CH1.61O0.61N0.17S0.01P0.03. The microbial analysis revealed Candidatus Kuenenia stuttgartiensis anammox bacteria and Nitrosomonas-like AOB were the two most dominant bacteria with 27.6% and 10.5% abundance, respectively, both of which formed spatially syntrophic co-immobilization within the micro-granules. The ex-situ activity tests showed the activity of NOB was well limited through DO regulation in the reactor. These results provide an alternative PN/A process configuration for low-strength wastewater treatment by sustaining microstate granules. Optimization of the nitrogen sludge loading rate and DO regulation are important for the successful performance.