Specific Ammonium Oxidation Rate Research Articles

Sludge enrichment and intermittent gradient aeration: Modulating microbial communities for start-up of partial nitrification in a sequencing batch reactor (SBR)

A novel approach has been proposed integrating sludge enrichment with intermittent gradient aeration to achieve partial nitrification (PN). Results indicated that this method suppressed nitrite-oxidizing bacteria (NOB) activity while maintained ammonia-oxidizing bacteria (AOB) activity, achieving an 82.87 % nitrite accumulation rate (NAR) during startup. During stable operation, specific ammonia oxidation rate and specific nitrite oxidation rate reached 18.44 and 2.67 mg N/g MLVSS/h, respectively. Shortening anoxic time from 30 to 20 min enhanced ammonia removal efficiency by 12.5 %, increasing NAR to 89.55 %. Further reduction to 5 min yielded 38.46 % and 91.88 %, respectively. AOB abundance enriched from 0.62 % to 22.28 %, with the AOB-to-NOB ratio jumping from 0.22 to 59.58. This research presented a rapid and effective strategy for achieving PN.

Biological treatment of N-methylpyrrolidone, cyclopentanone, and diethylene glycol monobutyl ether distilled residues and their effects on nitrogen removal in a full-scale wastewater treatment plant

Manufacturing processes in semiconductor and photonics industries involve the use of a significant amount of organic solvents. Recycle and reuse of these solvents produce distillate residues and require treatment before being discharged. This study aimed to evaluate the performance of the biological treatment system in a full-scale wastewater treatment plant that treats wastewater containing distillate residues from the recycling of electronic chemicals. Batch experiments were conducted to investigate the optimal operational conditions for the full-scale wastewater treatment plant. To achieve good nitrogen removal efficiency with effluent ammonia and nitrate concentrations below 20 mg N/L and 50 mg N/L, respectively, it was suggested to control the ammonia concentration and pH of the influent below 500 mg N/L and 8.0, respectively. In addition, the biodegradability of N-methylpyrrolidone, diethylene glycol monobutyl ether, and cyclopentanone distillate residues from the electronic chemicals manufacturing process were evaluated under aerobic, anoxic, and anaerobic conditions. N-methylpyrrolidone and cyclopentanone distillate residues were suggested to be treated under anoxic condition. However, substrate inhibition occurred when using cyclopentanone distillate residue as a carbon source with chemical oxygen demand (COD) levels higher than 866 mg/L and nitrate levels higher than 415 mg N/L. Under aerobic condition, the COD from both N-methylpyrrolidone and cyclopentanone distillate residues could be easily degraded. Nevertheless, a negative effect on nitrification was observed, with a prolonged lag time for ammonia oxidation as the initial COD concentration increased. The specific ammonia oxidation rate and nitrate production rate decreased under high COD concentration contributed by N-methylpyrrolidone and cyclopentanone distillate residues. Furthermore, the biodegradability of diethylene glycol monobutyl ether distillate residue was found to be low under aerobic, anoxic, and anaerobic conditions. With respect to the abundance of nitrogen removal microorganisms in the wastewater treatment plant, results showed that Comammox may have an advantage over ammonia oxidizing bacteria under high pH conditions. In addition, Comammox may have higher resistance to environmental changes. Dominance of Comammox over ammonia oxidizing bacteria under high ammonia condition was first reported in this study.

The superiority of hydrophilic polyurethane in comammox-dominant ammonia oxidation during low-strength wastewater treatment

Carriers have been extensively employed to enhance nitrification performance during low-strength wastewater treatment by retaining slow-growing ammonia oxidizing microorganisms (AOMs). Still, there is a dearth of systematic understanding of biofilm properties and microbial community structure formed on different carriers. In this study, hydrophilic polyurethane foam (PUF) carriers were prepared and compared with five widely used commercial carriers, namely Kaldness 3, Biochip, activated carbon, volcanic rock, and zeolite. The results indicated that the biofilms formed on carriers enhanced microbial ammonia oxidation activity. Additionally, the biofilm developed on the PUF demonstrated the most superior performance among all selected carriers, not only exhibiting the highest abundant and the most active AOMs, with amoA gene abundance of 1.41 × 1013 copies/m3 and specific ammonia oxidation rate of 9.84 g NH4+-N/(m3 × h), but also possessing a compact structure, with 3.41 kg VSS/m3 and 46.83 mg extracellular polymeric substances/g VSS. The high-throughput sequencing analysis revealed that the comammox (CMX) Nitrospira dominated on biofilm due to the intrinsically low apparent half-saturation constant for substrate. A unique ecological community structure was established on PUF, characterized by low species diversity and high homogeneity in alignment with community characteristics of CMX. The biofilms on PUF contributed to the proliferation of CMX Nitrospira dominated by Nitrospira nitrosa, achieving the highest proportion among colonial three AOMs at 86.58 %. The appropriate average pore size, superior hydrophilicity, and large specific surface area of PUF carriers provided a robust foundation for the exceptional ammonia oxidation performance of the formed biofilms.

Microplastics shaped performance, microbial ecology and community assembly in simultaneous nitrification, denitrification and phosphorus removal process

The widespread use of microplastics (MPs) has led to an increase in their discharge to wastewater treatment plants. However, the knowledge of impact of MPs on macro-performance and micro-ecology in simultaneous nitrification, denitrification, and phosphorus removal (SNDPR) systems is limited, hampering the understanding of potential risks posed by MPs. This study firstly comprehensively investigated the performance, species interactions, and community assembly under polystyrene (PS) and polyvinyl chloride (PVC) exposure in SNDPR systems. The results showed under PS (1, 10 mg/L) and PVC (1, 10 mg/L) exposure, total nitrogen removal was reduced by 3.38–10.15 %. PS and PVC restrained the specific rates of nitrite and nitrate reduction (SNIRR, SNRR), as well as the activities of nitrite and nitrate reductase enzymes (NIR, NR). The specific ammonia oxidation rate (SAOR) and activity of ammonia oxidase enzyme (AMO) were reduced only at 10 mg/L PVC. PS and PVC enhanced the size of co-occurrence networks, niche breadth, and number of key species while decreasing microbial cooperation by 5.85–13.48 %. Heterogeneous selection dominated microbial community assembly, and PS and PVC strengthened the contribution of stochastic processes. PICRUSt prediction further revealed some important pathways were blocked by PS and PVC. Together, the reduced TN removal under PS and PVC exposure can be attributed to the inhibition of SAOR, SNRR, and SNIRR, the restrained activities of NIR, NR, and AMO, the changes in species interactions and community assembly mechanisms, and the suppression of some essential metabolic pathways. This paper offers a new perspective on comprehending the effects of MPs on SNDPR systems.

Impact of salinity and organic matter on the ammonia-oxidizing archaea and bacteria in treating hypersaline industrial wastewater: amoA gene abundance and ammonia removal contributions.

Studies publishedrecently proposed that ammonia-oxidizing archaea (AOA) may bebeneficial for hypersaline (salinity > 50g NaCl L-1) industrial wastewater treatment. However, knowledge of AOA activity in hypersaline bioreactors is limited. This study investigated the effects of salinity, organic matter, and practical pickled mustard tuber wastewater (PMTW) on AOA and ammonia-oxidizing bacteria (AOB) in two sequencing batch biofilm reactors (SBBRs). Results showed that despite observed salinity inhibition (p < 0.05), both AOA and AOB contributed to high ammonia removal efficiency at a salinity of 70 g NaCl L-1 in the two SBBRs. The ammonia removal efficiency of SBBR2 did not significantly differ from that of SBBR1 in the absence of organic matter (p > 0.05). Batch tests and quantitative real-time PCR (qPCR) reveal that salinity and organic matter inhibition resulted in a sharp decline in specific ammonia oxidation rates and amoA gene copy numbers of AOA and AOB (p < 0.05). AOA demonstrated higher abundance and more active ammonia oxidation activity in hypersaline and high organic matter environments. Salinity was positively correlated with the potential ammonia oxidation contribution of AOA (p < 0.05), resulting in a potential transition from AOB dominance to AOA dominance in SBBR1 as salinity levels rose. Moreover, autochthonous AOA in PMTW promoted the abundance and ammonia oxidation activities of AOA in SBBR2, further elevating the nitrification removal efficiency after feeding the practical PMTW. AOA demonstrates greater tolerance tothe challenging hypersaline environment, making it a valuable candidate for the treatment of practical industrial wastewater with high salinity and organic content.

Short-term influence of polytetrafluoroethylene micro/nano-plastics on the inhibition of copper and/or ciprofloxacin on the nitrifying sludge activities based on concentration addition and independent action models

Short-term influence of polytetrafluoroethylene micro/nano-plastics (PTFE-MPs/NPs) on the inhibition of copper (Cu2+) and/or ciprofloxacin (CIP) on the nitrifying sludge activities was explored based on concentration addition (CA) and independent action (IA) models. The half maximal inhibitory concentration (IC50) of Cu2+, CIP, PTFE-MPs (3 μm), and PTFE-NPs (800 nm) on the specific ammonium oxidation rate (SAOR) of nitrifying sludge was 64.57, 51.29, 102.33 and 93.33 mg L−1, respectively, while those on the specific nitrite oxidation rate (SNOR) of nitrifying sludge were 77.62, 32.36, 104.70 and 97.72 mg L−1, respectively. Among the five binary mixtures and two ternary mixtures composed by Cu2+, CIP, and/or PTFE-MPs/NPs, it was found that the two joint inhibitory actions from ternary mixtures on the SAOR and SNOR of the sludge showed time-dependent characteristics by analyzing of CA and IA models, while the five combined inhibitory effects from different binary mixtures did not all have time-dependent features. The two joint inhibition actions from diverse ternary mixtures on the SAOR at the exposure time of 60 min and on the SNOR at 90 min showed always concentration-dependent features, while the combined inhibitions with concentration-dependent characteristics had never been observed in the binary Cu2+ and PTFE-NPs mixtures at different exposure time. The Cu2+, CIP, and PTFE-MPs mixtures (or Cu2+, CIP, and PTFE-NPs mixtures) had synergistic actions on the SAOR at 90 min and antagonistic effects on the SNOR at 60 min based on CA and IA models, and these combined inhibitions did not exhibit concentration-dependent characteristics. In contrast, the joint inhibitory effects (on the SAOR and SNOR) with concentration-dependent features were found in the binary mixtures of CIP and PTFE-MPs at different exposure time, and the join inhibition changed from synergism to antagonism as the increasing concentration of mixed CIP and PTFE-MPs. This study provides novel perspectives for understanding the combined influence of plastic particles with different sizes, antibiotics, and heavy metals on the biological wastewater treatment process.

Achieving partial nitrification: A strategy for washing NOB out under high DO condition

This study investigated the effect of high DO concentrations on PN. The experimental setup involved operating at high DO concentrations (1.5–2.5 mg/L) and environmental temperatures (15–20 °C) over a period of 180 days. Through a sludge enrichment process, the kinetic parameters of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were determined. Surprisingly, contrary to conventional reports, it was observed that NOB exhibited a stronger affinity for DO compared to AOB. As a result, high DO concentrations were necessary to provide favorable conditions for the growth of AOB. In order to establish PN, strategies including intermittent aeration, free ammonia (FA), and controlled sludge retention time (SRT) were employed. The successful PN was achieved with a specific ammonia oxidation rate of 24 mg N/g MLVSS/h and a specific nitrite oxidation rate below 0.10 mg N/g MLVSS/h. The nitrite accumulation rate (NAR) was maintained at 100% during stable operation. The abundance of Nitrosomonas, a typical genus of AOB, was found to be 68.62%, which surpasses previous studies in similar research. A slightly higher DO concentrations may increase energy consumption, but achieve higher efficiency and stability in PN. This study provided new insights into the application of PN in wastewater treatment.

Impacts of Polylactic Acid Microplastics on Performance and Microbial Dynamics in Activated Sludge System

A large number of microplastics (MPs) have been found in various stages of wastewater treatment plants, which may affect the functional microbial activity in activated sludge and lead to unstable pollutant removal performance. In this study, the effects of different concentrations of polylactic acid microplastics (PLA MPs) on system performance, nitrification and phosphorus (P) removal activities, and extracellular polymeric substances (EPS) were evaluated. The results showed that under the same influent conditions, low concentrations (50 particles/(g TS)) of PLA MPs had no significant effect on effluent quality. The average removal efficiencies of chemical oxygen demand, phosphate, and ammonia were all above 80%, and the average removal efficiencies of total nitrogen remained above 70%. High concentrations (200 particles/(g TS)) of PLA MPs inhibited the activities of polyphosphate-accumulating organisms (PAOs) and nitrifying bacteria. The specific anaerobic P release rate decreased from 37.7 to 23.1 mg P/(g VSS·h), and the specific aerobic P uptake rate also significantly decreased. The specific ammonia oxidation rate decreased from 0.67 to 0.34 mg N/(g VSS·h), while the change in the specific nitrite oxidation rate was not significant. The dosing of PLA MPs decreased the total EPS and humic acid content. As the concentration of PLA MPs increased, microbial community diversity increased. The relative abundance of potential PAOs (i.e., Acinetobacter) increased from 0.08 to 12.57%, while the relative abundance of glycogen-accumulating organisms (i.e., Competibacter and Defluviicoccus) showed no significant changes, which would lead to improved P removal performance. The relative abundance of denitrifying bacteria (i.e., Pseudomonas) decreased from 95.43 to 58.98%, potentially contributing to the decline in denitrification performance.

Specific ammonium oxidation and denitrification rates in an MBR treating real textile wastewater for simultaneous carbon and nitrogen removal

AbstractBACKGROUNDDissolved oxygen (DO) and aeration on/off time are the determining parameters for simultaneous carbon and nitrogen removal in biological treatment processes. The most effective way to decide the optimum operational conditions, e.g. aeration on/off time or DO concentration during the aeration period, is to determine specific nitrification/denitrification rates. For this purpose, the effects of DO (6 and 3 mg L−1) and aeration on/off time (from 2/2 to 90/360 min) on the specific ammonium oxidation and denitritation/denitrification rates in a membrane bioreactor (MBR) treating real textile wastewater were deeply investigated.RESULTSThe highest specific ammonium oxidation, denitritation and denitrification rates were obtained as 5.4, 3.8, and 5.3 mg N g−1 volatile suspended solids h−1, respectively, at an aeration on/off time of 90/360 min, which corresponded to the increase in specific ammonium oxidation rates by 1.8 and 2.1 times compared to continuous aeration conditions where DO was 6 and 3 mg L−1, respectively.CONCLUSIONHigher specific ammonium oxidation and denitrification rates can be achieved with the intermittent aeration compared to continuous aeration. Hence existing treatment plants can be retrofitted for higher performance with reduced energy requirements. © 2023 The Authors. Journal of Chemical Technology and Biotechnology published by John Wiley &amp; Sons Ltd on behalf of Society of Chemical Industry (SCI).

Effect of Cd(II) shock loading on performance, microbial enzymatic activity and microbial community in a sequencing batch reactor

The performance, microbial enzymatic activity and microbial community of a sequencing batch reactor (SBR) were explored under instantaneous Cd(II) shock loading. After a 24-h Cd(II) shock loading of 100 mg/L, the chemical oxygen demand and NH4+-N removal efficiencies decreased significantly from 92.73% and 99.56% on day 22 to 32.73% and 43% on day 24, respectively, and then recovered to the normal values gradually. The specific oxygen utilization rate (SOUR), specific ammonia oxidation rate (SAOR), specific nitrite oxidation rate (SNOR), specific nitrite reduction rate (SNIRR) and specific nitrate reduction rate (SNRR) decreased by 64.81%, 73.28%, 77.77%, 56.84% and 52.46% on day 23 in comparison with the absence of Cd(II) shock loading, respectively, and they gradually returned to the normal levels. The changing trends of their associated microbial enzymatic activities including dehydrogenase, ammonia monooxygenase, nitrite oxidoreductase, nitrite reductase and nitrate reductase were in accordance with SOUR, SAOR, SNOR, SNIRR and SNRR, respectively. Cd(II) shock loading promoted the microbial reactive oxygen species production and lactate dehydrogenase release, indicating that instantaneous shock caused oxidative stress and damaged to cell membranes of the activated sludge. The microbial richness and diversity, and the relative abundance of Nitrosomonas and Thauera obviously decreased under the stress of Cd(II) shock loading. PICRUSt prediction showed that Cd (II) shock loading significantly affected Amino acid biosynthesis, Nucleoside and nucleotide biosynthesis. The present results are conducive to take adequate precautions to reduce the adverse effect on bioreactor performance in wastewater treatment systems.

Dissolved oxygen benefits N-decanoyl-homoserine lactone regulated biological nitrogen removal system to resist acute ZnO nanoparticle exposure

The beneficial effects of N-decanoyl-homoserine lactone (C10-HSL), one of the typical N-acyl-homoserine lactones on biological nitrogen removal (BNR) system to resist the acute exposure of zinc oxide nanoparticles (ZnO NPs) has attracted extensive attentions. Nevertheless, the potential impact of dissolved oxygen (DO) concentration on the regulatory capacity of C10-HSL in the BNR system has yet to be investigated. This study conducted a systematic investigation of the impact of DO concentration on the C10-HSL-regulated BNR system against short-term ZnO NP exposure. Based on the findings, sufficient DO played a crucial role to improve the BNR system's resistance capacity to ZnO NPs. Under the micro-aerobic condition (0.5 mg/L DO), the BNR system was more sensitive to ZnO NPs. The ZnO NPs induced increased intracellular reactive oxygen species (ROS) accumulation, reduced antioxidant enzyme activities, and decreased specific ammonia oxidation rates in the BNR system. Furthermore, the exogenous C10-HSL had a positive effect on the BNR system's resistance to ZnO NP-induced stress, primarily by decreasing ZnO NPs-induced ROS generation and improving ammonia monooxygenase activities, especially under low DO concentrations. The findings contributed to the theoretical foundation for regulation strategy development of wastewater treatment plants under NP shock threat.

Long-term exposure of polytetrafluoroethylene-nanoplastics on the nitrogen removal and extracellular polymeric substances in sequencing batch reactor

The impact of polytetrafluoroethylene-nanoplastics (PTFE-NPs) on biological sewage disposal was delved, containing nitrogen remotion, microbiological activity and composition of extracellular polymer (EPS). The addition of PTFE-NPs reduced the removal efficiencies of chemical oxygen demand (COD) and ammonia nitrogen (NH4+-N) by 3.43 % and 2.35 %, respectively. In comparison with no PTFE-NPs, the specific oxygen uptake rate (SOUR), specific ammonia oxidation rate (SAOR), specific nitrite oxidation rate (SNOR) and specific nitrate reduction rate (SNRR) decreased by 65.26 %, 65.24 %, 41.77 % and 54.56 %, respectively. The PTFE-NPs inhibited the activities of nitrobacteria and denitrobacteria. It was worth noting that, nitrite oxidized bacterium was more resistant to adverse environments than ammonia oxidizing bacterium. Compared with no PTFE-NPs, the reactive oxygen species (ROS) content and lactate dehydrogenase (LDH) grew by 130 % and 50 % under PTFE-NPs pressure. The appearance of PTFE-NPs affected the normal function of microorganisms by inducing endocellular oxidative stress and destroying the completeness of the cytomembrane. The protein (PN) and polysaccharide (PS) levels in loosely bound EPS (LB-EPS) and tightly bound EPS (TB -EPS) increased by 4.96, 0.70, 3.07 and 0.71 mg g−1 VSS, under PTFE-NPs. Meanwhile, the PN/PS ratios of LB-EPS and TB -EPS increased from 6.18 and 6.41–11.04 and 9.29, respectively. The LB-EPS might provide sufficient binding sites for PTFE-NPs adsorption due to its loose and porous structure. The defense mechanism of bacteria against PTFE-NPs was mainly the PN in loosely bound EPS. Moreover, the functional groups referred to the complexation of EPS with PTFE-NPs were mainly related to N-H, CO, and C-N in proteins and O-H in polysaccharides.

Efficient partial nitrification with hybrid nitrifying granular sludge based on a simultaneous fill/draw SBR mode

In this study, a simultaneous fill/draw SBR was applied to investigate the feasibility of partial nitrification process with inoculation of matured aerobic granular sludge. The system operated stably over 120 days with the relatively high ammonium removal efficiency (≥ 98.83%) and nitrite accumulation rate (≥ 89.60%). Moreover, a hybrid flocs/granules system was formed stably after long-term operation. The nitrite-oxidizing bacteria (NOB) was suppressed effectively because of the combined effect of simultaneous fill/draw mode and intermittent aeration conditions. Furthermore, batch tests were separately tested with isolated granules (> 200 μm) and flocs (< 200 μm), showing that the specific ammonia oxidation rate of granules and flocs were 15.94 ± 2.85 and 66.77 ± 0.83 mg N/(g MLSS·h), respectively. Correspondingly, the abundance of Nitrosomonas as a typical AOB in granules (6.24%) and flocs (11.94%) was obtained via the microbial diversity analysis, while NOB was almost hardly detected in granules and flocs.

An enriched ammonia-oxidizing microbiota enables high removal efficiency of ammonia in antibiotic production wastewater

High ammonia concentration hinders the efficient treatment of antibiotic production wastewater (APW). Developing effective ammonia oxidation wastewater treatment strategies is an ideal approach for facilitating APW treatment. Compared with traditional nitrification strategies, the partial nitrification process is more eco-friendly, less energy-intensive, and less excess sludge. The primary limiting factor of the partial nitrification process is increasing ammonia-oxidizing bacteria (AOB) while decreasing nitrite-oxidizing bacteria (NOB). In this study, an efficient AOB microbiota (named AF2) was obtained via enrichment of an aerobic activated sludge (AS0) collected from a pharmaceutical wastewater treatment plant. After a 52-day enrichment of AS0 in 250 mL flasks, the microbiota AE1 with 69.18% Nitrosomonas microorganisms was obtained. Subsequent scaled-up cultivation in a 10 L fermenter led to the AF2 microbiota with 59.22% Nitrosomonas. Low concentration of free ammonia (FA, < 42.01 mg L−1) had a negligible effect on the activity of AF2, and the nitrite-nitrogen accumulation rate (NAR) of AF2 was 98% when FA concentration was 42.01 mg L−1. The specific ammonia oxidation rates (SAORs) at 30 °C and 15 °C were 3.64 kg NH4+-N·kg MLVSS−1·d−1 and 1.43 kg NH4+-N·kg MLVSS−1·d−1 (MLVSS: mixed liquor volatile suspended solids). The SAOR was 0.52 kg NH4+-N·kg MLVSS−1·d−1 when the NaCl concentration was increased from 0 to 20 g L−1, showing that AF2 functioning was stable in a high-level salt environment. The ammonia oxidation performance of AF2 was verified by treating abamectin and lincomycin production wastewater. The NARs of AF2 used for abamectin and lincomycin production wastewater treatment were >90% and the SAORs were 2.39 kg NH4+-N·kg MLVSS−1·d−1 and 0.54 kg NH4+-N·kg MLVSS−1·d−1, respectively, which was higher than the traditional biological denitrification process. In summary, AF2 was effective for APW treatment via enhanced ammonia removal efficiency, demonstrating great potential for future industrial wastewater treatment.

Transient disturbance of CeO2 nanoparticles enhances N2O emissions during biological wastewater treatment

Although the negative effects of CeO2 nanoparticles (CeO2 NPs) on wastewater treatment processes have been confirmed, their impacts on the greenhouse gas nitrous oxide (N2O) emissions during biological nitrogen removal (BNR) remain unclear. Herein, the response of BNR and associated N2O emissions in a sequencing batch reactor (SBR) to different loads (1–50 mg/L) of CeO2 NPs was investigated. Although CeO2 NPs induced the dose-dependent inhibition on NH4+ oxidation rate, the NH4+-N and total nitrogen (TN) removal efficiencies did not change. The N2O emission flux was enhanced along with the increase in the CeO2 NP dose, and there was a positive correlation between N2O emission and nitrite accumulation. In addition, NH4+ oxidation activity indicators ammonia monooxygenase (AMO) activity, specific ammonia oxidation rate (SAOR), and specific oxygen uptake rate (SOUR) were suppressed under the 25 and 50 mg/L CeO2 NP stresses. The increase in reactive oxygen species (ROS) levels induced by NP stress resulted in the downregulation of the amoA and nosZ gene expression. Furthermore, the CeO2 NP shock load caused the severe suppression of NH4+ oxidizing bacteria-related Nitrosomonas activities while promoted the denitrifying glycogen-accumulating organism (potential N2O producer)-related Dechloromonas activities. In summary, this study proves evidence that the transient disturbance of CeO2 NPs would negatively affect the biological nitrogen transformation to release more N2O.

Nitrogen removal characteristics of biofilms in each area of a full-scale AAO oxidation ditch process.

Plastic carriers were installed in different areas of a full-scale anaerobic/anoxic/aerobic (AAO) oxidation ditch process, and the dynamics of nitrogen removal, biofilm morphologies, and microorganism species were investigated. The results showed that the biofilm at the front of the aerobic area (dissolved oxygen [DO]=0.93mgL-1) provided the best denitrification, with specific nitrate and nitrite reduction rates of 10.16 and 3.78mg·(g·h)-1, respectively. The biofilm in the middle of the aerobic area (DO=1.27mgL-1) exhibited the best nitrification performance, with a maximum specific ammonia oxidation rate of 3.21mg·(g·h)-1. The biofilm at the end of the aerobic area (DO=0.01mgL-1) exhibited the highest anammox potential with a maximum specific anammox rate of 0.67mg·(g·h)-1. No correlation was observed between the specific nitrogen removal rates and abundance of nitrogen-removing microorganisms at the genus level. Biofilm denitrification during the process was primarily completed by heterotrophic denitrifying bacteria (Thauera, Acinetobacter, Hyphomicrobium, and Thermomonas). Aerobic denitrifying bacteria (0.19% Thauera and 0.34% Hyphomicrobium) were identified as the main denitrifying bacteria in the middle of the aerobic area. The dominant nitrifying microorganisms in the middle of the aerobic area were Nitrosomonas (0.50%) and Nitrospira (1.04%). A biofilm in the end of the aerobic area exhibited specific anammox potential, which may have been related to the dominance of 0.024% Candidatus Brocadia. Kinetic analysis revealed that adding plastic carriers to the front and middle of the aerobic area contributed to stable nitrogen removal efficiency.

Achieving PN through the selective recovery of AOB activity in inactivated nitrifying bacteria: Combined aerobic starvation and FA

A novel strategy combining aerobic starvation and free ammonia (FA) was proposed to achieve partial nitrification (PN). The impact of the combined strategy on nitrifying bacteria was explored in a 200-day experiment. The effluent concentration of ammonia was below the detection limits (0.1 mg/L), and the effluent concentration of nitrite and nitrate was 68.12 mg/L and 3.46 mg/L without adding carbon source to the artificial wastewater. The nitrite accumulation rate (NAR) was maintained at 90.15% even when the dissolved oxygen (DO) concentration was 1.50 mg/L. Further analysis showed that PN was achieved by selectively restoring the activity of ammonia-oxidizing bacteria (AOB) in nitrifying bacteria that had lost their activity after starvation. The specific ammonia oxidation rate (SAOR) was 46.25 mg N/g MLVSS/h, and the specific nitrate product rate (SNPR) was only 0.73 mg N/g MLVSS/h in the stable operation stage. The increase in AOB abundance (from 2.79% to 7.13%) and the decrease in nitrite-oxidizing bacteria (NOB) abundance (from 8.75% to 1.44%) explained this phenomenon. Finally, the analyses on the secretion of extracellular polymer substance (EPS), strategies to resist harsh environments, and physical properties of sludge explored the potential mechanism and provided references for applying the combined strategy.

Exploration of mechanisms for calcium phosphate precipitation and accumulation in nitrifying granules by investigating the size effects of granules

Calcium phosphate could be accumulated in aerobic granules, which attracts attention recently for phosphorus removal and recovery from wastewater. In this study, partial nitrifying granules with high calcium precipitate content were sorted into different size groups for characterization and evaluation to reveal the dynamic balance of granules at stead state and relevant calcium phosphate precipitation and accumulation mechanism. It was found that light yellow small granules without calcium precipitates but high microbial activity co-existed with deep brown granules with calcium precipitate of around 91% and low microbial activity. Characterization with specific oxygen uptake rates, specific ammonium oxidation rates, calcium and phosphate removal rates from solution, EPS contents, elemental compositions by energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and scanning electron microscopy (SEM) for different size groups of granules enabled a proposal of a new hypothesized mechanism for calcium precipitation and accumulation. With this proposed mechanism, it is believed that sufficient granule retention time in reactors was critical for the accumulation of calcium precipitates followed by a slow microbial growth rate of biomass due to mass transfer resistance. The co-precipitation of calcium carbonate and calcium phosphate mainly occurred in granules with a size less than 710 µm while calcium phosphate dominant minerals were accumulated in granules larger than 710 µm. The results and conclusions in this study shed light on the mechanisms of calcium phosphate accumulation in granules, which could be used to better operate and control aerobic granular sludge with calcium phosphates for phosphorus removal and recovery.

Size dependent impacts of a model microplastic on nitrification induced by interaction with nitrifying bacteria

Two sizes of polystyrene (PS) were compared to investigate their impact on nitrification. The smaller PS (50 nm) had a higher impact than the larger PS (500 nm). Lower NO2- and NO3- accumulation was observed in the 50 nm PS treatment. There was no significant difference in DIN concentration between the control and 500 nm PS treatments. PS treatment did not have a significant influence on the specific ammonia oxidation rate, but the specific nitrite utilization rate was the lowest in the 50 nm PS treatment. The changes in transcript levels of amoA gene did not correspond well with the observed changes in DIN concentrations, suggesting that the effects of 50 nm PS treatment might be unrelated to biological phenomena, for which an actual uptake of PS is needed. The fluorescent images revealed that the smaller PS can easily access bacterial cells, which corroborated the results of inhibition of nitrification by the smaller PS. Notably, most of the PS particles did not penetrate bacterial cells, suggesting that the observed effects of 50 nm PS on nitrification might be due to disruption of the membrane potential of the cells.

Hydroxyapatite Precipitation and Accumulation in Granules and Its Effects on Activity and Stability of Partial Nitrifying Granules at Moderate and High Temperatures

Precipitation and accumulation of calcium phosphate in granular sludge has attracted research attention recently for phosphate removal and recovery from wastewater. This study investigated calcium phosphate accumulation from granulation stage to steady state by forming heterotrophic granules at different COD/N ratios at 21 and 32 °C, respectively, followed by the transformation of heterotrophic granules to partial nitrifying granules. It was found that mature granules accumulated around 60–80% minerals in granules, much higher than young granules with only around 30% ash contents. In addition, high temperature promoted co-precipitation of hydroxyapatite and calcite in granules with more calcite than hydroxyapatite and only 4.1% P content, while mainly hydroxyapatite was accumulated at the moderate temperature with 7.7% P content. The accumulation of minerals in granules at the high temperature with 75–80% ash content also led to the disintegration and instability of granules. Specific ammonium oxidation rates were reduced, as well, from day 58 to day 121 at both temperatures due to increased mineral contents. These results are meaningful to control or manipulate granular sludge for phosphorus removal and recovery by forming and accumulating hydroxyapatite in granules, as well as for the maintenance of microbial activities of granules.