Anammox-Mediated Hydroxyapatite Granules: Physicochemical Properties, 3D Hierarchy, and Biofilm Thickness.

Biomineralization inspired the development of simultaneous biological transformations and chemical precipitation for simultaneous nitrogen removal and phosphorus recovery from wastewater, which could compensate for the incapacity of phosphorus management in the new biological route of anaerobic ammonium oxidation (anammox). In this study, we strengthened anammox-mediated biomineralization by long-term feeding of concentrated N, P, and Ca substrates, and a self-assembled matrix of anammox bacteria and hydroxyapatite (HAP) was fabricated in a granular shape, defined as HAP-anammox granules. HAP was identified as the dominant mineral using elemental analysis, X-ray diffraction, and Raman spectroscopy. The intensive precipitation of HAP resulted in a higher inorganic fraction and substantially improved settleability of anammox biomass, which facilitated HAP precipitation by acting as nucleation and metabolically elevated pH. By using X-ray microcomputed tomography, we visually represented the hybrid texture of interwoven HAP pellets and biomass, the core-shell layered architecture of different-sized HAP-anammox granules, and their homogeneously regulated thickness of the outer biofilm (from 118 to 635 µm). This unique architecture endows HAP-anammox granules with outstanding settleability, active biofilm, and tightly bonded biofilm with the carrier, which may explain the excellent performance of these HAP-anammox granules under various challenging operational conditions in previous studies.

Improved Properties and Enhancement Strategies of Hydroxyapatite-Based Functional Granular Sludge for a High-Rate Partial Nitritation/Anammox System.

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.

Towards advanced simultaneous nitrogen removal and phosphorus recovery from digestion effluent based on anammox-hydroxyapatite (HAP) process: Focusing on a solution perspective.

In this paper, the state-of-the-art information on the anammox-HAP process is summarized. The mechanism of this process is systematically expounded, the enhancement of anammox retention by HAP precipitation and the upgrade of phosphorus recovery by anammox process are clarified. However, this process still faces several challenges, especially how to deal with the âˆ¼ 11% nitrogen residues and to purify the recovered HAP. For the first time, an anaerobic fermentation (AF) combined with partial denitrification (PD) and anammox-HAP (AF-PD-Anammox-HAP) process is proposed to overcome the challenges. By AF of the organic impurities of the anammox-HAP granular sludge, organic acid is produced to be used as carbon source for PD to remove the nitrogen residues. Simultaneously, pH of the solution drops, which promotes the dissolution of some inorganic purities such as CaCO3. In this way, not only the inorganic impurities are removed, but the inorganic carbon is supplied for anammox bacteria.

Anammox upflow hybrid reactor: Nitrogen removal performance and potential for phosphorus recovery.

Echoing to the call of recovering high-value-added chemicals from wastewater and achieving carbon-neutral operation in wastewater treatment, an anammox upflow hybrid reactor was successfully applied for nitrogen removal, and the potential for phosphorus recovery was put forward. Moreover, the spatial pattern of removal capacities, and distribution of biomass and HAP precipitates were recognized and demonstrated as height-oriented. The intensity of HAP precipitates was highly consistent with the amount of anammox biomass and the relative abundance of the Candidatus Kuenenia, indicating that HAP formation was encouraged by the anammox reaction itself and heterogeneous nucleation induced by organic matters (proteins and polysaccharides). The fixed bed also played an important role in immobilizing the anammox biomass, secreted organic matrix, and HAP precipitates. This finding also provoked the thought that in the anammox process, HAP precipitation was more achievable, effective and practicable using the fixed-carrier system.

The phosphorus harvest from low-temperature mainstream wastewater through iron phosphate crystallization in a pilot-scale partial nitritation/anammox reactor.

The phosphorus harvest along nitrogen removal in the partial nitritation/anammox (PNA) reactor is promising for saving space and simplifying the management of mainstream wastewater treatment facilities. In this study, the phosphorus recovery from the low-temperature mainstream wastewater was explored through iron phosphate crystallization in a pilot-scale PNA reactor. With the COD-alleviated municipal wastewater as the influent, the ammonium concentration of about 50 mg/L and the phosphorus concentration ranged from 5.4 to 7.1 mg/L, under the temperature of 15 °C and the addition of external ferrous iron of 14 mg/L, the achieved nitrogen removal efficiency and the phosphorus removal efficiency were 37.6 % and 62.7 %, respectively. The good settleability of sludge indicated that the formed iron phosphate was well combined with the biomass. The quantitative analysis confirmed that the main iron phosphate in dry sludge was graftonite, and qualitative analysis confirmed that the equivalent of P2O5 content in the sludge was 5.8 %, which was suitable as fertilizer on agricultural land to realize the direct recycle of discharged phosphorus. In all, this study proposed a pioneering scheme to realize the nitrogen removal and phosphorus cycle in human society and given a meaningful reference for further research and application.

Material mass balance and elemental flow analysis in a submerged anaerobic membrane bioreactor for municipal wastewater treatment towards low-carbon operation and resource recovery.

The anaerobic membrane bioreactor (AnMBR) has gained huge attention as a municipal wastewater (MWW) treatment process that combined high organics removal, a low sludge yield and bioenergy recovery. In this study, a 20 L AnMBR was set up and operated steadily for 70 days in temperate conditions with an HRT of 6 h and a flux of 12 LMH for the treatment of real MWW, focusing on the behavior of the major elements (C, N, P and S) from an elemental balance perspective. The results showed that the AnMBR achieved more than 85 % COD removal, a low sludge yield (0.081 gVSS/gCODremoved) and high methane production (0.31 L-CH4/gCODremoved) close to the theoretical value. The elemental flow analysis revealed that the AnMBR converted 77 % of the influent COD to methane (57 % gaseous and 20 % dissolved) and 6 % of the COD for sludge production. In addition, the AnMBR converted 34 % of the total carbon to energy-generated carbon, and only 3 % was in the form of CO2 in the biogas for further upgradation, which was in line with the concept of carbon neutrality. Since little nitrogen or phosphorus were removed, the permeate was nutrient-rich and further treatment to recover the nutrients would be required. This study illustrates the superior performance of the AnMBR for MWW treatment with a microscopic view of elemental behavior and provides a reference for implementing the mainstream AnMBR process in carbon-neutral wastewater treatment plants.

Biofilm growth characterization and treatment performance in a single stage partial nitritation/anammox process with a biofilm carrier.

Biofilm carriers can avoid microorganism washout while maintaining a high amount of biomass, but are also associated with a long biofilm formation period and biofilm aging. A single stage partial nitritation/anammox process (single stage PN/A) reactor was setup to study the biofilm growth characterization and treatment performance under an NLR of 0.53 to 0.90 gN/L/d over one year. Biofilm growth was divided into three stages: the formation stage, maturation stage and aging stage. The initial biofilm was observed at day 84. A nitrogen removal efficiency of 83.4% was achieved at an NLR of 0.90 gN/L/d during the mature biofilm stage. Starvation, nitrogen gas accumulation and hydroxyapatite formation resulted in biofilm aging. After mechanical stirring treatment, biofilm reactivation was achieved by biofilm re-formation within one month. There is clear potential for phosphorus recovery, as indicated by the 5.24% - 6.29% phosphorus content in the biofilm (similar to the 5%-7% phosphorus content in enhanced biological phosphate removal sludge). The AnAOB genera abundance in the biofilm maintained at a high level of 18.25%-32.31%, while the abundance of AnAOB increased from the initial 4.10% to 13.78% after mechanical stirring treatment in the suspended sludge ensured biofilm reactivation. The results of this study clearly show that mechanical stirring treatment can be used to achieve the biofilm reactivation as the biofilm fills with the hollow cylindrical carrier. This study has potential as a useful reference for the realization of the wide application of the biofilm single stage PN/A process in the future.

Granular activated carbon promoting re-granulation of anammox-hydroxyapatite granules for stable nitrogen removal at low phosphate concentration.

Anaerobic ammonium oxidation (anammox) coupled with hydroxyapatite (HAP) crystallization not only achieves simultaneous nitrogen removal and phosphorus recovery, but also cultivates excellent anammox granules. However, a floatation and wash-out of anammox-HAP granules was occurred at low phosphate concentrations. In this study, a reactor inoculated with mature anammox-HAP granules and fed with low phosphate (5 mg P/L) was added with granular activated carbon (GAC) to maintain sludge granulation and nitrogen-removing stability. At influent total nitrogen >800 mg/L and nitrogen loading rate ~ 9.8 kg/m3/d, a satisfactory nitrogen removal of around 88% was maintained during 140 days of operation. Insufficient phosphate supplement resulted in a sludge bulking, with suspended solid and sludge density decreased whereas sludge water content and expansion ratio increased due to HAP loss. Nevertheless, the sludge re-granulation was found at the later stage as the proportion of granules in 2.8- 3.35 mm went up to 37.4% after large granules disintegrated into small pieces at the initial stage. The settling velocity was finally ranging from 129.8 to 182.2 m/h. In addition, Candidatus Brocadia was increased from 2.1% to 20.1% and dominated in the microbial community. These findings suggest GAC was able to promote re-granulation of anammox-HAP granules at low phosphate concentration, which avoids sludge flotation and widens their application as an inoculum.

Microstructure and granulation cycle mechanisms of anammox-HAP coupled granule in the anammox EGSB reactor.

The formation of anammox-hydroxyapatite (HAP) coupled granules has been shown to be an approach to efficient nitrogen removal and phosphorus recovery in the anammox EGSB reactor. However, the granulation cycle mechanism of anammox-HAP coupled granules for sustainable regeneration and growth is still not well understood. In this study, the microstructure, chemical composition and microbial structure of a total of six different-sized granules, from 0.25 mm to 2.8 mm, was determined. An SEM-EDS analysis indicated that the small granules (<0.5 mm) were composed of poly-pellet clusters with anammox biofilms attached to the HAP cores, and the large granules (>0.5 mm) consisted of a three-layer structure: a surface anammox biofilm layer, a middle connection layer, and a HAP mineral inner core. The analysis of elemental composition and microbial structure suggested homogenous granular characteristics regardless of granule size. The dominant microorganisms were anammox bacteria of Candidatus Kuenenia stuttgartiensis and heterotrophic denitrifying bacteria. Based on these results, a granulation cycle mechanism for anammox-HAP coupled granules was proposed for the first time. The growth of the small granules with the simultaneous enlargement of anammox biofilms and HAP cores results in the formation of large granules. Large granules regenerate new small granules in a two-step procedure. The first step is the separation of embryo HAP crystals from the mother core via heterogeneous growth, and the second step is the separation of the biofilms due to biodegradation and shear stress.

Enhancing phosphorus recovery from sewage sludge using anaerobic-based processes: Current status and perspectives.

Anaerobic-based processes are green and sustainable technologies for phosphorus (P) recovery from sewage sludges economically and are promising in practical application. However, the P release efficiency is always not satisfied. In this paper, the P release mechanisms (regarding to different P species) from sewage sludge using anaerobic-based processes are systematically summarized. The obstacles of P release and the updated achievements of enhancing P release from sewage sludges are analyzed and discussed. It can be concluded that different P species can release from sewage sludge via different anaerobic-based processes. Extracellular polymeric substances and excessive metal ions are the two main limiting factors to P release. Acid fermentation and anaerobic fermentation with sulfate reduction could be two promising ways, with P release efficiencies of up to 64% and 63%. Based on the summarization and discussion, perspectives on practical application of P recovery from sewage sludge using anaerobic-based processes are proposed.

The impact of calcium supplementation on methane fermentation and ammonia inhibition of fish processing wastewater.

The effect of trace metal supplementation on the methane fermentation of fish processing wastewater (FPW) was studied in both batch and continuous experiments using a self-agitated anaerobic baffled reactor (SA-ABR). In the batch experiments, a single supplementation of Ca2+, Co2+ and Fe2+ was show to have a significant positive impact on the performance of methane fermentation. The continuous experiment results showed that supplementation with 1.5 g-Ca2+/L-substrate remarkably enhanced the performance of methane fermentation of the SA-ABR in treating FPW with the optimal organic loading rate achieved at 7.62 g-COD/L/d. During the steady states (stages 2 to 5), the average removal efficiencies of COD, protein, carbohydrate and lipid were 89, 85, 80 and 91%, respectively. The biogas conversion rates were in the range of 0.39 to 0.45 L-biogas/g-COD with a high methane content of 74%. Besides, Ca2+ supplementation also improved the resistance of the methane fermentation system to ammonia inhibition.

Improvement of coffee grounds high solid thermophilic methane fermentation by co-digestion with in-situ produced waste activated sludge: Performance and stability.

The feasibility of in-situ stabilization in the co-digestion of coffee grounds (CG) and waste activated sludge (WAS) was investigated. Two lab-scale thermophilic continuous stirred tank reactors (CSTR), R1 and R2 were operated with substrates that contained different WAS ratios, S1 (WAS% = 20%) and S2 (WAS% = 30%). During the whole process, there was no external supply of ammonia and trace elements. The volatile solid (VS) removal efficiency of R1 and R2 was comparable, and the biogas yield of R1 (0.467 ± 0.100 L/g-VSin) was slightly higher than R2 (0.408 ± 0.020 L/g-VSin). The total ammonia nitrogen (TAN) of R1 and R2 was 482 ± 32 and 884 ± 24 mg/L, respectively. The stoichiometry formulas of co-digestion were established to calculate the theoretical microbial yield coefficients and the requirements of microorganism reproduction. A comparison between the theoretical requirements and experimental values showed that co-digestion with WAS could avoid supply for an external supply of minerals. For the net energy production, R1 and R2 could generate 6342 and 5069 kWh of electricity daily, respectively.

Hydroxyapatite crystallization-based phosphorus recovery coupling with the nitrogen removal through partial nitritation/anammox in a single reactor.

For digestion effluent treatment, while the anammox-based process has been successfully applied for nitrogen removal, in most cases, phosphorus (P) represents another major concern. In this study, a novel process, integrating the partial nitritation/anammox and hydroxyapatite crystallization (PNA-HAP) in a single airlift reactor, was developed for the simultaneous nitrogen removal and P recovery from synthetic digestion effluent. With a stable influent P concentration of 20.0 mg/L, an HRT of 6 h, and alternating increases of influent calcium and ammonium, the final achieved nitrogen removal rate was 1.2 kg/m3/d and the P removal efficiency was 83.0%. The settleability of sludge was desirably enhanced with the calcium addition and a high biomass concentration was achieved in reactor. Quantitative and qualitative analyses confirmed that HAP was the main inorganic content in sludge, which could be harvested for P recovery. According to the Scanning Electron Microscope observation and the Energy Dispersive X-ray spectrometry analysis, the microbes were mainly distributed on the outer layer of the sludge aggregate, while the HAP mainly in the interior. The relevant theoretical calculation also revealed that the sludge discharge manipulation has direct effect on the sludge composition and aggregate structure. In sum, the results are evidence of the feasibility of simultaneous nitrogen removal and P recovery through one-stage PNA-HAP process for digestion effluent.

Simultaneous nitrogen removal and phosphorus recovery using an anammox expanded reactor operated at 25 °C.

While anammox is a cost-effective nitrogen treatment process for wastewater with high nutrient strength, phosphorus remains untouched during this process and needs further treatment. In this study, the nitrogen removal and phosphorus recovery were achieved simultaneously at 25 °C using an anammox expanded bed. A stable high nitrogen removal efficiency of 83.7 ± 4.8% at a 1500 mgN/L influent total nitrogen concentration and a phosphorus removal efficiency of 94.2 ± 1.2% at 100 mg P/L influent total phosphorus were obtained during continuous operation. The effects of the nitrogen loading rate, hydraulic retention time (HRT), pH, Ca2+ and PO43- concentration on the phosphorus removal was verified in the long-term operation of the reactor. The sludge produced contained a high content of phosphorus mainly in the form of hydroxyapatite (HAP), and the sludge composition strongly reflected the nitrogen and phosphorus loading. The structure of the anammox-HAP composite granules was illustrated by the use of fluorescence in situ hybridization (FISH) and Raman spectroscopic mapping analysis. The results in this study indicate that by controlling the operation parameters, it is possible to integrate a high efficiency phosphorus recovery with the anammox process, and significantly reduce the nutrient loading for further wastewater treatment.

Towards more efficient nitrogen removal and phosphorus recovery from digestion effluent: Latest developments in the anammox-based process from the application perspective.

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.

Effect of additional food waste slurry generated by mesophilic acidogenic fermentation on nutrient removal and sludge properties during wastewater treatment.

Fermentation slurry from food waste (FSFW) generated by acidogenic fermentation at mesophilic temperature was utilized to improve the nutrients removal from wastewater. Organic acids (such as lactate and volatile fatty acids) in the FSFW behaved as readily biodegradable carbon sources, while the particulate and macromolecular organics acted as slowly biodegradable carbon sources during denitrification processes. The FSFW dosage significantly influenced the nitrogen removal performance, and a C/N ratio (in terms of chemical oxygen demand to nitrogen ratio) of 8 could achieve complete denitrification in the batch tests. In a sequencing batch reactor (SBR) using FSFW for long-term wastewater treatment, extracellular polymeric substances (EPS) gradually accumulated, sludge particle size significantly increased, and microbial communities were selectively enriched, which contributed to promoting the nitrogen (>80%) and phosphate (90.1%) removal efficiencies. Overall, the FSFW produced by acidogenic fermentation under mesophilic temperature served as an excellent intermediary between FW valorization and wastewater treatment.

Carbon source and phosphorus recovery from iron-enhanced primary sludge via anaerobic fermentation and sulfate reduction: Performance and future application.

Anaerobic fermentation and sulfate reduction (AF-SR) was firstly used for recovery of carbon sources and phosphorus from Fe-enhanced primary sludge (Fe-sludge). With FeCl3 dosage of 30 mg Fe/L, 63.0% of the chemical oxygen demand (COD) and 97.3% of the phosphorus were concentrated from sewage into Fe-sludge. Batch anaerobic fermentation tests of Fe-sludge with and without sulfate addition (AF-SR and control) were performed. The results showed that volatile fatty acid concentrations of the control and AF-SR were 211.0 and 270.2 mg COD/g volatile suspended solids, respectively. Furthermore, 33.2% (control) and 56.2% (AF-SR) of the total phosphorus in Fe-sludge was released. The recovery performances of carbon source and phosphorus were calculated based on struvite precipitation. The available carbon source of the AF-SR system was 44.5% higher than that of the control. A novel integrated wastewater and sludge treatment process based on chemically enhanced primary sedimentation and AF-SR is proposed for future application.

Process optimization of anammox-driven hydroxyapatite crystallization for simultaneous nitrogen removal and phosphorus recovery.

Based on the requirements for advanced treatment and resource recovery of nitrogen and phosphorus pollutants in wastewater, the coupled anammox and hydroxyapatite crystallization (anammox-HAP) process was studied with an aim of achieving high efficiency and low energy consumption during simultaneous nitrogen and phosphorus removal. In the long-term experiments and batch tests, the effects of substrate conditions (nitrogen and phosphorus load, calcium concentration, etc.) on the nitrogen removal and phosphorus recovery efficiencies were investigated. The granular structure and crystal properties were analyzed together with microscopic characterization methods, and the formation mechanism of coupled anammox-HAP granules was verified. Based on these experiments, a theoretical model and technical method for realizing the coupled process were established, and a reference for practical engineering application was provided.

Energy recovery potential of thermophilic high-solids co-digestion of coffee processing wastewater and waste activated sludge by anaerobic membrane bioreactor.

A large amount of wastewater is generated in the processing of coffee from fruit to cup. Thermophilic high-solids co-digestion of coffee processing wastewater (CPW) and waste activated sludge (WAS) has been succeeded by anaerobic membrane bioreactor (AnMBR). Chemical oxygen demand (COD) removal efficiencies of 92 ±â€¯3% with an average methane yield of 0.28 LCH4/gCODremoved were achieved at a high solids content of 50 g/L in the AnMBR. The optimal digestion performance of 82.4% removal COD conversion to CH4 was achieved at hydraulic retention time (HRT) 10 d. Energy balance analysis revealed AnMBR has succeeded in energy positive at all the HRTs. The net energy potential (NEP) was determined to average 2.12-2.82 kJ/gCOD, amongst which the maximum NEP was achieved at HRT 15 d. These results indicated the high-solids co-digestion by AnMBR is a promising approach to maximize the bioenergy conversion from the co-substrate of CPW and WAS.

Nutrients removal performance and sludge properties using anaerobic fermentation slurry from food waste as an external carbon source for wastewater treatment.

Enhancement of nitrogen and phosphate removal using thermophilic fermentation slurry from food waste (FSFW) as external carbon source was investigated. Based on the batch tests, the soluble and particulate fractions of the FSFW acted as easily and slowly biodegradable carbon sources, respectively, and the fermented slurry showed the combined nutrients removal properties of soluble and solid organics. During the long-term operation of a sequencing batch reactor (SBR) with FSFW for wastewater treatment, the sludge particle size increased obviously, the bacterial metabolic capacity improved significantly, and some functional microorganisms were enriched selectively, which significantly promoted the nitrogen removal efficiency (approximately 90%) by enhancing the anoxic denitrification and simultaneous nitrification and denitrification (SND) processes. Moreover, high phosphate removal efficiency (above 98%) was achieved through the aerobic and anoxic phosphate accumulation processes. Thus, using the FSFW as supplementary carbon source is a suitable solution for both food waste disposal and wastewater treatment.