A review on anammox process for the treatment of antibiotic-containing wastewater: Linking effects with corresponding mechanisms

• Anammox is promising for nitrogen removal from antibiotic-containing wastewater. • Most antibiotics could inhibit the anammox performance and activity. • Antibiotic pressure promoted the increase in antibiotic resistance genes (ARGs). • Antibiotic-resistance mechanisms of anammox bacteria are speculated. Antibiotic is widely present in the effluent from livestock husbandry and the pharmaceutical industry. Antibiotics in wastewater usually have high biological toxicity and even promote the occurrence and transmission of antibiotic resistant bacteria and antibiotic resistance genes. Moreover, most antibiotic-containing wastewater contains high concentration of ammonia nitrogen. Improper treatment will lead to high risk to the surrounding environment and even human health. The anaerobic ammonium oxidation (anammox) with great economic benefit and good treatment effect is a promising process to remove nitrogen from antibiotic-containing wastewater. However, antibiotic inhibition has been observed in anammox applications. Therefore, a comprehensive overview of the single and combined effects of various antibiotics on the anammox system is conducted in this review with a focus on nitrogen removal performance, sludge properties, microbial community, antibiotic resistance genes and anammox-involved functional genes. Additionally, the influencing mechanism of antibiotics on anammox consortia is summarized. Remaining problems and future research needs are also proposed based on the presented summary. This review provides a better understanding of the influences of antibiotics on anammox and offers a direction to remove nitrogen from antibiotic-containing wastewater by the anammox process.     

Simultaneous Feammox and anammox process facilitated by activated carbon as an electron shuttle for autotrophic biological nitrogen removal

• The autotrophic nitrogen removal combining Feammox and Anammox was achieved. • Activated carbon can be used as an electron shuttle to enhance Feammox activity. • Fe(III) was reduced to Fe(II) and the secondary Fe(II) mineral (FeOOH) was obtained. • The iron-reducing bacteria and Anammox consortium was enriched simultaneously. Ferric iron reduction coupled with anaerobic ammonium oxidation (Feammox) is a novel ferric-dependent autotrophic process for biological nitrogen removal (BNR) that has attracted increasing attention due to its low organic carbon requirement. However, extracellular electron transfer limits the nitrogen transformation rate. In this study, activated carbon (AC) was used as an electron shuttle and added into an integrated autotrophic BNR system consisting of Feammox and anammox processes. The nitrogen removal performance, nitrogen transformation pathways and microbial communities were investigated during 194 days of operation. During the stable operational period (days 126–194), the total nitrogen (TN) removal efficiency reached 82.9%±6.8% with a nitrogen removal rate of 0.46±0.04 kg-TN/m³/d. The contributions of the Feammox, anammox and heterotrophic denitrification pathways to TN loss accounted for 7.5%, 89.5% and 3.0%, respectively. Batch experiments showed that AC was more effective in accelerating the Feammox rate than the anammox rate. X-ray photoelectron spectroscopy (XPS) analyses showed the presence of ferric iron (Fe(III)) and ferrous iron (Fe(II)) in secondary minerals. X-ray diffraction (XRD) patterns indicated that secondary iron species were formed on the surface of iron-AC carrier (Fe/AC), and Fe(III) was primarily reduced by ammonium in the Feammox process. The phyla Anaerolineaceae (0.542%) and Candidatus Magasanikbacteria (0.147%) might contribute to the Feammox process, and Candidatus Jettenia (2.10%) and Candidatus Brocadia (1.18%) were the dominative anammox phyla in the bioreactor. Overall, the addition of AC provided an effective way to enhance the autotrophic BNR process by integrating Feammox and anammox.     

Biological conversion pathways of sulfate reduction ammonium oxidation in anammox consortia

• The SRAO phenomena tended to occur only under certain conditions. • High amount of biomass and non-anaerobic condition is requirement for SRAO. • Anammox bacteria cannot oxidize ammonium with sulfate as electron acceptor. • AOB and AnAOB are mainly responsible for ammonium conversion. • Heterotrophic sulfate reduction mainly contributed to sulfate conversion. For over two decades, sulfate reduction with ammonium oxidation (SRAO) had been reported from laboratory experiments. SRAO was considered an autotrophic process mediated by anammox bacteria, in which ammonium as electron donor was oxidized by the electron acceptor sulfate. This process had been attributed to observed transformations of nitrogenous and sulfurous compounds in natural environments. Results obtained differed largely for the conversion mole ratios (ammonium/sulfate), and even the intermediate and final products of sulfate reduction. Thus, the hypothesis of biological conversion pathways of ammonium and sulfate in anammox consortia is implausible. In this study, continuous reactor experiments (with working volume of 3.8L) and batch tests were conducted under normal anaerobic (0.2≤DO<0.5 mg/L) / strict anaerobic (DO<0.2 mg/L) conditions with different biomass proportions to verify the SRAO phenomena and identify possible pathways behind substrate conversion. Key findings were that SRAO occurred only in cases of high amounts of inoculant biomass under normal anaerobic condition, while absent under strict anaerobic conditions for same anammox consortia. Mass balance and stoichiometry were checked based on experimental results and the thermodynamics proposed by previous studies were critically discussed. Thus anammox bacteria do not possess the ability to oxidize ammonium with sulfate as electron acceptor and the assumed SRAO could, in fact, be a combination of aerobic ammonium oxidation, anammox and heterotrophic sulfate reduction processes.     

Sludge fermentation liquid addition attained advanced nitrogen removal in low C/N ratio municipal wastewater through short-cut nitrification-denitrification and partial anammox

• Sludge fermentation liquid addition resulted in a high NAR of 97.4%. • Extra NH₄⁺-N from SFL was removed by anammox in anoxic phase. • Nitrogen removal efficiency of 92.51% was achieved in municipal wastewater. • The novel system could efficiently treat low COD/N municipal wastewater. Biological nitrogen removal of wastewater with low COD/N ratio could be enhanced by the addition of wasted sludge fermentation liquid (SFL), but the performance is usually limited by the introducing ammonium. In this study, the process of using SFL was successfully improved by involving anammox process. Real municipal wastewater with a low C/N ratio of 2.8–3.4 was treated in a sequencing batch reactor (SBR). The SBR was operated under anaerobic-aerobic-anoxic (AOA) mode and excess SFL was added into the anoxic phase. Stable short-cut nitrification was achieved after 46d and then anammox sludge was inoculated. In the stable period, effluent total inorganic nitrogen (TIN) was less than 4.3 mg/L with removal efficiency of 92.3%. Further analysis suggests that anammox bacteria, mainly affiliated with Candidatus_Kuenenia, successfully reduced the external ammonia from the SFL and contributed approximately 28%–43% to TIN removal. Overall, this study suggests anammox could be combined with SFL addition, resulting in a stable enhanced nitrogen biological removal.     

Partial anammox achieved in full scale biofilm process for typical domestic wastewater treatment

• A full scale biofilm process was developed for typical domestic wastewater treatment. • The HRT was 8 h and secondary sedimentation tank was omitted. • Candidatus Brocadia were enriched in the HBR with an abundance of 2.89%. • Anammox enabled a stable ammonium removal of ~15% in the anoxic zone. The slow initiation of anammox for treating typical domestic wastewater and the relatively high footprint of wastewater treatment infrastructures are major concerns for practical wastewater treatment systems. Herein, a 300 m³/d hybrid biofilm reactor (HBR) process was developed and operated with a short hydraulic retention time (HRT) of 8 h. The analysis of the bacterial community demonstrated that anammox were enriched in the anoxic zone of the HBR process. The percentage abundance of Candidatus Brocadia in the total bacterial community of the anoxic zone increased from 0 at Day 1 to 0.33% at Day 130 and then to 2.89% at Day 213. Based upon the activity of anammox bacteria, the removal of ammonia nitrogen (NH₄⁺-N) in the anoxic zone was approximately 15%. This showed that the nitrogen transformation pathway was enhanced in the HBR system through partial anammox process in the anoxic zone. The final effluent contained 12 mg/L chemical oxygen demand (COD), 0.662 mg/L NH₄⁺-N, 7.2 mg/L total nitrogen (TN), and 6 mg/L SS, indicating the effectiveness of the HBR process for treating real domestic wastewater.     

Start-up of PN-anammox system under low inoculation quantity and its restoration after low-loading rate shock

• PN-A was start-up under low inoculation amount and a higher NRR was achieved. • PN-anammox system was successfully restored by aggressive sludge discharge. • Increase in granular sludge was the important factor to rapid recovery. • Enrichment of AOB and AnAOB in granular sludge favors the stable operation. Partial nitritation (PN)-anaerobic ammonium oxidation (anammox) is a promising pathway for the biological treatment of wastewater. However, the destruction of the system caused by excessive accumulation of nitrate in long-term operation remains a challenge. In this study, PN-anammox was initialized with low inoculation quantity in an air-lift reactor. The nitrogen removal rate of 0.71 kgN/(m³·d) was obtained, which was far higher than the seed sludge (0.3 kgN/(m³·d)). Thereafter, excess nitrate build-up was observed under low-loading conditions, and recovery strategies for the PN-anammox system were investigated. Experimental results suggest that increasing the nitrogen loading rate as well as the concentration of free ammonium failed to effectively suppress the nitrite oxidation bacteria (NOB) after the PN-anammox system was disrupted. Afterwards, effluent back-flow was added into the reactor to control the up-flow velocity. As a result, an aggressive discharge of sludge that promoted the synergetic growth of functional bacteria was achieved, leading to the successful restoration of the PN-anammox system. The partial nitritation and anammox activity were in balance, and an increase in nitrogen removal rate up to 1.07 kgN/(m³·d) was obtained with a nitrogen removal efficiency of 82.4% after recovery. Besides, the proportion of granular sludge (over 200 mm) increased from 33.67% to 82.82%. Ammonium oxidation bacteria (AOB) along with anammox bacteria were enriched in the granular sludge during the recovery period, which was crucial for the recovery and stable operation of the PN-anammox system.     

The main anammox-based processes,the involved microbes and the novel process concept from the application perspective

• The PNA, denitratation/anammox, and DAMO/anammox process are reviewed together. • Denitratation/anammox-based process is promising in mainstream treatment. • DAMO and denitratation processes realize the higher nitrogen removal efficiency. • The utilization of metabolism diversity of functional microbe is worth exploring. • An effective waste treatment system concept is proposed. Anammox technology has been widely researched over the past 40-year from the laboratory-scale to full-scale. It is well-known that in actual applications, the solo application of anammox is not feasible. Since both ammonium and nitrite are prerequisites based on the reaction mechanism, the pre-treatment of wastewater is necessary. With the combination of anammox process and other pre-treatment processes to treat the actual wastewater, many types of anammox-based processes have been developed with distinct nitrogen removal performance. Thus, in order to heighten the awareness of researchers to the developments and accelerate the application of these processes to the treatment of actual wastewater, the main anammox-based processes are reviewed in this paper. It includes the partial nitritation/anammox process, the denitratation/anammox (PD/A) process, the denitrifying anaerobic methane oxidation/anammox (DAMO/A) process, and more complex deuterogenic processes. These processes have made the breakthroughs in the application of the anammox technology, such as the combination of nitrification and PD/A process can achieve stability and reliability of nitrogen removal in the treatment of mainstream wastewater, the PD/A process and the DAMO/A have brought about further improvements in the total nitrogen removal efficiency of wastewater. The diversity of functional microbe characteristics under the specific condition indicate the wide application potential of anammox-based processes, and further exploration is necessary. A whole waste treatment system concept is proposed through the effective allocation of above mentioned processes, with the maximum recovery of energy and resources, and minimal environmental impact.     

Overlooked nitrogen-cycling microorganisms in biological wastewater treatment

• AOA and comammox bacteria can be more abundant and active than AOB/NOB at WWTPs. • Coupled DNRA/anammox and NO_x-DAMO/anammox/comammox processes are demonstrated. • Substrate level, SRT and stressors determine the niches of overlooked microbes. • Applications of overlooked microbes in enhancing nitrogen removal are promising. Nitrogen-cycling microorganisms play key roles at the intersection of microbiology and wastewater engineering. In addition to the well-studied ammonia oxidizing bacteria, nitrite oxidizing bacteria, heterotrophic denitrifiers, and anammox bacteria, there are some other N-cycling microorganisms that are less abundant but functionally important in wastewater nitrogen removal. These microbes include, but not limited to ammonia oxidizing archaea (AOA), complete ammonia oxidation (comammox) bacteria, dissimilatory nitrate reduction to ammonia (DNRA) bacteria, and nitrate/nitrite-dependent anaerobic methane oxidizing (NO_x-DAMO) microorganisms. In the past decade, the development of high-throughput molecular technologies has enabled the detection, quantification, and characterization of these minor populations. The aim of this review is therefore to synthesize the current knowledge on the distribution, ecological niche, and kinetic properties of these “overlooked” N-cycling microbes at wastewater treatment plants. Their potential applications in novel wastewater nitrogen removal processes are also discussed. A comprehensive understanding of these overlooked N-cycling microbes from microbiology, ecology, and engineering perspectives will facilitate the design and operation of more efficient and sustainable biological nitrogen removal processes.     

Characterization of CANON reactor performance and microbial community shifts with elevated COD/N ratios under a continuous aeration mode

COD/N at low ratios (0–0.82) improved N removals of CANON. CANON performance decreased after COD/N up to 0.82. The relative abundance of AOB decreased continuously with increasing COD/N. AOB outcompeted at a high COD load led to CANON failure. The relative abundance of AnAOB decreased and increased with increasing COD/N. The effects of increasing COD/N on nitrogen removal performance and microbial structure were investigated in a SBR adopting a completely autotrophic nitrogen removal over nitrite process with a continuous aeration mode (DO at approximately 0.15–0.2 mg/L). As the COD/N increased from 0.1 to≤0.59, the nitrogen removal efficiency (NRE) increased from 88.7% to 95.5%; while at COD/N ratios of 0.59–0.82, the NRE remained at 90.7%–95.5%. As the COD/N increased from 0.82 to 1.07, the NRE decreased continuously until reaching 60.1%. Nitrosomonas sp. (AOB) and Candidatus Jettenia (anammox bacteria) were the main functional genera in the SBR. As the COD/N increased from 0.10 to 1.07, the relative abundance of Nitrosomonas decreased from 13.4% to 2.0%, while that of Candidatus Jettenia decreased from 35% to 9.9% with COD/N<0.82 then increased to 45.4% at a COD/N of 1.07. Aerobic heterotrophic bacteria outcompeted AOB at high COD loadings (650 mg/L) because of oxygen competition, which ultimately led to deteriorated nitrogen removal performance.     

Effects of Fe(II) on anammox community activity and physiologic response

• 0.12 mmol/L Fe(II) enhanced the total anammox activity and bacterial abundance best. • 0.09 mmol/L Fe(II) led to the best performance on relative anammox activity. • 0.75 mmol/L Fe(II) had an immediate but recoverable inhibition on anammox activity. • More genes but not relative level were expressed at higher Fe(II) concentration. Though there are many literatures studying the effects of iron on anammox process, these studies only focus on the reactor performance and/or the microbial community changes, the detailed effects and mechanisms of Fe(II) on anammox bacterial activity and physiology have not been explored. In this study, four Fe(II) concentrations (0.03, 0.09, 0.12 and 0.75 mmol/L) were employed into the enriched anammox culture. The enhancement and inhibition effects of Fe(II) on anammox process and bacterial physiology were investigated. It was discovered that the anammox process and bacterial growth were enhanced by 0.09 and 0.12 mmol/L Fe(II), in which the 0.12 mmol/L Fe(II) had advantage in stimulating the total anammox activity and bacterial abundance, while 0.09 mmol/L Fe(II) enhanced the relative anammox activity better. The anammox activity could be inhibited by 0.75 mmol/L Fe(II) immediately, while the inhibition was recoverable. Both 0.09 and 0.12 mmol/L Fe(II) induced more genes being expressed, while didn’t show a stimulation on the relative expression level of functional genes. And anammox bacteria showed a stress response to detoxify the Fe inhibition once inhibited by 0.75 mmol/L Fe(II). This study provides more information about physiologic response of anammox bacteria to external influence (enhancement and inhibition), and may also instruct the future application of anammox process in treating various sources of wastewater (containing external disturbances such as heavy metals) and/or different treatment strategies (e.g. from side-stream to main-stream).     

Genome-resolved metagenomic analysis reveals different functional potentials of multiple Candidatus Brocadia species in a full-scale swine wastewater treatment system

● Four Ca. Brocadia species were observed during the spontaneously enrichment. ● Novel anammox species SW510 and SW773 dominated the full-scale ecosystem. ● Urease and cyanase genes were detected in the new anammox genomes. ● Functional differentiation potentially facilitated co-occurrence of anammox species. The increasing application of anammox processes suggests their enormous potential for nitrogen removal in wastewater treatment facilities. However, the functional potentials and ecological differentiation of cooccurring anammox species in complex ecosystems have not been well elucidated. Herein, by utilizing functional reconstruction and comparative genome analysis, we deciphered the cooccurring mechanisms of four Candidatus Brocadia species that were spontaneously enriched in a full-scale swine wastewater treatment system. Phylogenetic analysis indicated that species SW172 and SW745 were closely related to Ca. Brocadia caroliniensis and Ca. Brocadia sapporoensis, respectively, whereas the dominant species SW510 and SW773, with a total average abundance of 34.1%, were classified as novel species of the genus Ca. Brocadia. Functional reconstruction revealed that the novel species SW510 can encode both cytochrome cd₁-type nitrite reductase and hydroxylamine oxidase for nitrite reduction. In contrast, the detected respiratory pentaheme cytochrome c nitrite reductase and acetate kinase genes suggested that SW773 likely reduced nitrite to ammonium with acetate as a carbon source. Intriguingly, the presence of genes encoding urease and cyanase indicated that both novel species can use diverse organic nitrogen compounds in addition to ammonia and nitrite as substrates. Taken together, the recovery and comparative analysis of these anammox genomes expand our understanding of the functional differentiation and cooccurrence of the genus Ca. Brocadia in wastewater treatment systems.     

Insights into simultaneous anammox and denitrification system with short-term pyridine exposure: Process capability,inhibition kinetics and metabolic pathways

• Short-term effect of the pyridine exposure on the SAD process was investigated. • The SAA at 150 mg/L pyridine reduced by 56.7% of the maximum value. • Inhibition kinetics models and inhibitory parameters were indicated. • Collaboration of AnAOB, HDB and PDB promoted the SAD. • Possible metabolic pathways of nitrogen and pyridine were proposed. In-depth knowledge on the role of pyridine as a bottleneck restricting the successful application of anammox-based process treating refractory coking wastewater remains unknown. In this study, the effect of short-term pyridine addition on a simultaneous anammox and denitrification (SAD) system fed with 25–150 mg/L pyridine was explored. The short-term operation showed that the highest total nitrogen (TN) removal efficiency was achieved at 25–50 mg/L of pyridine. As the pyridine addition increased, the contribution of the anammox pathway in nitrogen removal decreased from 99.3% to 79.1%, while the denitrification capability gradually improved. The specific anammox activity (SAA) at 150 mg/L pyridine decreased by 56.7% of the maximum SAA. The modified non-competitive inhibition model indicated that the 50% inhibitory concentration (IC₅₀) of pyridine on anammox was 84.18 mg/L and the substrate inhibition constant (K_i) of pyridine for self-degradation was 135.19 mg/L according to the Haldane model. Moreover, high-throughput sequencing confirmed the abundance of Candidatus Kuenenia as the amount of anammox species decreased, while the amounts of denitrifiers and pyridine degraders significantly increased as the pyridine stress increased. Finally, the possible pathways of nitrogen bioconversion and pyridine biodegradation in the SAD system were elucidated through metagenomic analysis and gas chromatography/mass spectrometry results. The findings of this study enlarge the understanding of the removal mechanisms of complex nitrogenous pyridine-containing wastewater treated by the SAD process.     

Research progress and prospects of complete ammonia oxidizing bacteria in wastewater treatment

• Comammox bacteria have unique physiological characteristics. • Comammox bacteria are widely distributed in natural and artificial systems. • Comammox bacteria have the potential to reduce N₂O emissions. • Coupling comammox bacteria with DEAMOX can be promoted in wastewater treatment. • Comammox bacteria have significant potential for enhancing total nitrogen removal. Complete ammonia oxidizing bacteria, or comammox bacteria (CAOB), can oxidize ammonium to nitrate on its own. Its discovery revolutionized our understanding of biological nitrification, and its distribution in both natural and artificial systems has enabled a reevaluation of the relative contribution of microorganisms to the nitrogen cycle. Its wide distribution, adaptation to oligotrophic medium, and diverse metabolic pathways, means extensive research on CAOB and its application in water treatment can be promoted. Furthermore, the energy-saving characteristics of high oxygen affinity and low sludge production may also become frontier directions for wastewater treatment. This paper provides an overview of the discovery and environmental distribution of CAOB, as well as the physiological characteristics of the microorganisms, such as nutrient medium, environmental factors, enzymes, and metabolism, focusing on future research and the application of CAOB in wastewater treatment. Further research should be carried out on the physiological characteristics of CAOB, to analyze its ecological niche and impact factors, and explore its application potential in wastewater treatment nitrogen cycle improvement.     

Methanation and chemolitrophic nitrogen removal by an anaerobic membrane bioreactor coupled partial nitrification and Anammox

● Efficient carbon methanation and nitrogen removal was achieved in AnMBR-PN/A system. ● AOB outcompeted NOB in PN section by limiting aeration and shortening SRT. ● The moderate residual organic matter of PN section triggered PD in anammox unit. ● AnAOB located at the bottom of UASB played an important role in nitrogen removal. An AnMBR-PN/A system was developed for mainstream sewage treatment. To verify the efficient methanation and subsequent chemolitrophic nitrogen removal, a long-term experiment and analysis of microbial activity were carried out. AnMBR performance was less affected by the change of hydraulic retention time (HRT), which could provide a stable influent for subsequent PN/A units. The COD removal efficiency of AnMBR was > 93% during the experiment, 85.5% of COD could be recovered in form of CH₄. With the HRT of PN/A being shortened from 10 to 6 h, nitrogen removal efficiency (NRE) of PN/A increased from 60.5% to 80.4%, but decreased to 68.8% when the HRT_PN/A further decreased to 4 h. Microbial analysis revealed that the highest specific ammonia oxidation activity (SAOA) and the ratio of SAOA to specific nitrate oxidation activity (SNOA) provide stable NO₂⁻-N/NH₄⁺-N for anammox, and anammox bacteria (mainly identified as Candidatus Brocadia) enriched at the bottom of Anammox-UASB might play an important role in nitrogen removal. In addition, the decrease of COD in Anammox-UASB indicated partial denitrification occurred, which jointly promoted nitrogen removal with anammox.     

Reutilize tire in microbial fuel cell for enhancing the nitrogen removal of the anammox process coupled with iron-carbon micro-electrolysis

• MFC promoted the nitrogen removal of anammox with Fe-C micro-electrolysis. • Reutilize pyrolysis waste tire as micro-electrolysis and electrode materials. • Total nitrogen removal efficiency of modified MFC increased to 85.00%. • Candidatus kuenenia and SM1A02 were major genera responsible for nitrogen removal. In this study, microbial fuel cells (MFCs) were explored to promote the nitrogen removal performance of combined anaerobic ammonium oxidation (anammox) and Fe-C micro-electrolysis (CAE) systems. The average total nitrogen (TN) removal efficiency of the modified MFC system was 85.00%, while that of the anammox system was 62.16%. Additionally, the effective operation time of this system increased from six (CAE system alone) to over 50 days, significantly promoting TN removal. The enhanced performance could be attributed to the electron transferred from the anode to the cathode, which aided in reducing nitrate/nitrite in denitrification. The H⁺ released through the proton exchange membrane caused a decrease in the pH, facilitating Fe corrosion. The pyrolyzed waste tire used as the cathode could immobilize microorganisms, enhance electron transport, and produce a natural Fe-C micro-electrolysis system. According to the microbial community analysis, Candidatus kuenenia was the major genus involved in the anammox process. Furthermore, the SM1A02 genus exhibited the highest abundance and was enriched the fastest, and could be a novel potential strain that aids the anammox process.     

Taxonomic and functional variations in the microbial community during the upgrade process of a full-scale landfill leachate treatment plant – from conventional to partial nitrification-denitrification

• Upgrade process was investigated in a full-scale landfill leachate treatment plant. • The optimization of DO can technically achieve the shift from CND to PND process. • Nitrosomonas was mainly responsible for ammonium oxidation in PND system. • An obviously enrichment of Thauera was found in the PND process. • Enhanced metabolic potentials on organics was found during the process update. Because of the low access to biodegradable organic substances used for denitrification, the partial nitrification-denitrification process has been considered as a low-cost, sustainable alternative for landfill leachate treatment. In this study, the process upgrade from conventional to partial nitrification-denitrification was comprehensively investigated in a full-scale landfill leachate treatment plant (LLTP). The partial nitrification-denitrification system was successfully achieved through the optimizing dissolved oxygen and the external carbon source, with effluent nitrogen concentrations lower than 150 mg/L. Moreover, the upgrading process facilitated the enrichment of Nitrosomonas (abundance increased from 0.4% to 3.3%), which was also evidenced by increased abundance of amoA/B/C genes carried by Nitrosomonas. Although Nitrospira (accounting for 0.1%–0.6%) was found to stably exist in the reactor tank, considerable nitrite accumulation occurred in the reactor (reaching 98.8 mg/L), indicating high-efficiency of the partial nitrification process. Moreover, the abundance of Thauera, the dominant denitrifying bacteria responsible for nitrite reduction, gradually increased from 0.60% to 5.52% during the upgrade process. This process caused great changes in the microbial community, inducing continuous succession of heterotrophic bacteria accompanied by enhanced metabolic potentials toward organic substances. The results obtained in this study advanced our understanding of the operation of a partial nitrification-denitrification system and provided a technical case for the upgrade of currently existing full-scale LLTPs.     

Enhancing the efficiency of nitrogen removing bacterial population to a wide range of C:N ratio (1.5:1 to 14:1) for simultaneous C & N removal

• Simultaneous C & N removal in Methammox occurs at wide C:N ratio. • Biological Nitrogen Removal at wide C:N ratio of 1.5:1 to 14:1 is not reported. • Ammonia removal shifted from mixotrophy to heterotrophy at high C:N ratio. • Acetogenic population compensated for ammonia oxidizers at high C:N ratio. • Methanogens increase the plasticity of nitrogen removers at high C:N ratio. High C:N ratio in the wastewater limits biological nitrogen removal (BNR), especially in anammox based technologies. The present study attempts to improve the COD tolerance of the BNR process by associating methanogens with nitrogen removing bacterial (NRB) populations. The new microbial system coined as ‘Methammox’, was investigated for simultaneous removal of COD (C) and ammonia (N) at C:N ratio 1.5:1 to 14:1. The ammonia removal rate (11.5 mg N/g VSS/d) and the COD removal rates (70.6 mg O/g VSS/d) of Methammox was close to that of the NRB (11.1 mg N/g VSS/d) and the methanogenic populations (77.9 mg O/g VSS/d), respectively. The activities established that these two populations existed simultaneously and independently in ‘Methammox’. Further studies in biofilm reactor fetched a balanced COD and ammonia removal (55%–60%) at a low C:N ratio (≤2:1) and high C:N ratio (≥9:1). The population abundance of methanogens was reasonably constant, but the nitrogen removal shifted from mixotrophy to heterotrophy as the C:N ratio shifted from low (C:N≤2:1) to high (C:N≥9:1). The reduced autotrophic NRB (ammonia- and nitrite-oxidizing bacteria and Anammox) population at a high C:N ratio was compensated by the fermentative group that could carry out denitrification heterotrophically. The functional plasticity of the Methammox system to adjust to a broad C:N ratio opens new frontiers in biological nitrogen removal of high COD containing wastewaters.     

The impact of ultrasonic treatment on activity of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria in activated sludge

Conditions for ultrasonic treatment to achieve partial nitritation are optimized. Ultrasound reduces metabolic activity and releases intracellular metabolites. Mechanical shearing is essential to inhibit nitrite oxidation. The ultrasonic treatment of sludge has been considered as an effective method to facilitate the partial nitritation of municipal sewage. This study aims to reveal the effects of ultrasound on ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The impact factors including ultrasonic irradiation time and intensity, sludge concentration, thermal effect and released free radicals were studied. The maximized difference between the changes in AOB and NOB activities were obtained with 10 g mixed liquor suspended solids (MLSS)/L, using 0.9 kJ/mL ultrasonic energy density and 12 h interval time. The increased ultrasonic intensity destroyed the floc structure of activated sludge, increased the microbial death, and decreased the cellular ATP level. Further, the mechanism exploration indicated that the mechanical shearing could be a critical factor in achieving the nitritation with inhibitory effect on nitrite oxidation.     

Distribution and removal of antibiotic resistance genes during anaerobic sludge digestion with alkaline,thermal hydrolysis and ultrasonic pretreatments

Sludge digestion is critical to control the spread of ARGs from wastewater to soil. Fate of ARGs in three pretreatment-AD processes was investigated. UP was more efficient for ARGs removal than AP and THP in pretreatment-AD process. The total ARGs concentration showed significant correlation with 16S rRNA gene. The bacteria carrying ARGs could be mainly affiliated with Proteobacteria. Sewage sludge in the wastewater treatment plants contains considerable amount of antibiotic resistance genes (ARGs). A few studies have reported that anaerobic digestion (AD) could successfully remove some ARGs from sewage sludge, but information on the fate of ARGs in sludge pretreatment-AD process is still very limited. In this study, three sludge pretreatment methods, including alkaline, thermal hydrolysis and ultrasonic pretreatments, were compared to investigate the distribution and removal of ARGs in the sludge pretreatment-AD process. Results showed that the ARGs removal efficiency of AD itself was approximately 50.77%, and if these three sludge pretreatments were applied, the total ARGs removal efficiency of the whole pretreatment-AD process could be improved up to 52.50%–75.07%. The ultrasonic pretreatment was more efficient than alkaline and thermal hydrolysis pretreatments. Although thermal hydrolysis reduced ARGs obviously, the total ARGs rebounded considerably after inoculation and were only removed slightly in the subsequent AD process. Furthermore, it was found that the total ARGs concentration significantly correlated with the amount of 16S rRNA gene during the pretreatment and AD processes, and the bacteria carrying ARGs could be mainly affiliated with Proteobacteria.