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The treatment of antibiotic wastewater, which is known for its micro-toxicity, inhibition, and poor biochemistry, poses significant challenges, including complex processes, high energy demands, and secondary pollution. Bio-Fenton, a novel Fenton technology, enables the in situ production of H2O2 at near-neutral pH, having low energy requirements and sustainable properties, and reduces the hazards of H2O2 transportation and storage. We preliminary self-designed a heterogeneous Bio-Fenton reactor. An aerobic SBBR system with pure algae, pure bacteria, and bacteria-algae symbiosis was first constructed to investigate the optimal process conditions through the effects of carbon source concentration, light duration, bamboo charcoal filling rate, and dissolved oxygen (DO) content on the H2O2 production and COD removal. Second, the reactor was constructed by adding iron-carrying catalysts to remove ROX and SDZ wastewater. The results demonstrated that the optimal operating parameters of aerobic SBBR were an influent carbon source concentration of 500 mg/L, a water temperature of 20 ± 2 °C, pH = 7.5, a dissolved oxygen content of 5 mg/L, a light-dark ratio of 12 h:12 h, a light intensity of 2500 Lux, an HRT of 10 h, and a bamboo charcoal filling rate of 33%. Given these conditions, the bacterial-algal system was comprehensively found to be the most suitable biosystem for this experiment. Ultimately, the dynamically coupled Bio-Fenton process succeeded in the preliminary removal of 41.32% and 42.22% of the ROX and SDZ from wastewater, respectively.

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Sequencing batch biofilm reactor (SBBR) has the potential to treat hypersaline high-strength nitrogen wastewater by simultaneous nitrification-denitrification (SND). Dissolved oxygen (DO) and aeration modes are major factors affecting pollutant removal. Low DO (0.35-3.5 mg/L) and alternative anoxic/aerobic (A/O) mode are commonly used for municipal wastewater treatment, however, the appropriate DO concentration and operation mode are still unknown under hypersaline environment because of the restricted oxygen transfer in denser extracellular polymeric substances (EPS) barrier and the decreased carbon source consumption during the anoxic phase. Herein, two SBBRs (R1, fully aerobic mode; R2, A/O mode) were used for the treatment of hypersaline high-strength nitrogen wastewater (200 mg/L NH4+-N, COD/N of 3 and 3% salinity). The results showed that the relatively low DO (2 mg/L) could not realize effective nitrification, while high DO (4.5 mg/L) evidently increased nitrification efficiency by enhancing oxygen transfer in denser biofilm that was stimulated by high salinity. A stable SND was reached 16 days faster with a ∼10% increase of TN removal under A/O mode. Mechanism analysis found that denser biofilm with coccus and bacillus were present in A/O mode instead of filamentous microorganisms, with the secretion of more EPS. Corynebacterium and Halomonas were the dominant genera in both SBBRs, and HN-AD process might assist partial nitrification-denitrification (PND) for highly efficient TN removal in biofilm systems. By using the appropriate operation mode and parameters, the average NH4+-N and TN removal efficiency could respectively reach 100% and 70.8% under the NLR of 0.2 kg N·m-3·d-1 (COD/N of 3), which was the highest among the published works using SND-based SBBRs in treatment of saline high-strength ammonia nitrogen (low COD/N) wastewater. This study provided new insights in biofilm under hypersaline stress and provided a solution for the treatment of hypersaline high-strength nitrogen (low COD/N) water.

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Domestic wastewater source-separated treatment has attracted wide attention due to the efficiency improvement of sewage treatment systems, energy saving, resource reuse, and the construction and operation cost saving of pipeline networks. Nonetheless, the excess source-separated urine still demands further harmless treatment. Sequencing batch biofilm reactor (SBBR), a new type of composite biofilm reactor developed by filling different fillers into the sequential batch reactor (SBR) reactor, has higher pollutant removal performance and simpler operation and maintenance. However, the phosphorus removal ability of the SBBR filling with conventional fillers is still limited and needs further improvement. In this study, we developed two new fillers, the self-fabricated filler A and B (SFA/SFB), and compared their source-separated urine treatment performance. Long-term treatment experimental results demonstrated that the SBBR systems with different fillers had good removal performance on the COD and TN in the influent, and the removal rate increased with the increasing HRT. However, only the SBBR system with the SFA showed excellent PO43--P and TP removal performance, with the removal rates being 83.7 ± 11.9% and 77.3 ± 13.7% when the HRT was 1 d. Microbial community analysis results indicated that no special bacteria with strong phosphorus removal ability were present on the surface of the SFA. Adsorption experimental results suggested that the SFA had better adsorption performance for phosphorus than the SFB, but it could not always have stronger phosphorus adsorption and removal performance during long-term operation due to the adsorption saturation. Through a series of characterizations such as SEM, XRD, and BET, it was found that the SFA had a looser structure due to the use of different binder and production processes, and the magnesium in the SFA gradually released and reacted with PO43- and NH4+ in the source-separated urine to form dittmarite and struvite, thus achieving efficient phosphorus removal. This study provides a feasible manner for the efficient treatment of source-separated urine using the SBBR system with self-fabricated fillers.

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With the aim of addressing the difficult problem of biodegradable organic nitrogen in biochemical effluent of a printing and dyeing industrial park, the combined ozonation-sequencing batch biofilm reactor (O3-SBBR) process was used for advanced treatment. The influencing factors and degradation kinetics were analyzed; quenching experiments were carried out; and the types of free radicals, succinate dehydrogenase activity, and denitrification function genes were determined. The results showed that the suitable ozonation condition was pH 8.0-8.5, O3 concentration was approximately 35.0 mg·L-1, O3 dosage was approximately 100.0 mg·L-1, and reaction time was 90.0-120.0 min. Organic nitrogen in the biochemical effluent by ozonation conformed to the pseudo first-order kinetic model, and the maximum rate constant k was 0.01035 min-1 (experimental conditions:pH 8.0, ozone dosage 150.0 mg·L-1, and ozone concentration 35.0 mg·L-1). Ozonation significantly improved the denitrification performance of the sequencing biofilm batch reactor (SBBR), and the denitrification efficiency increased from 19.8% (SBBR) to 32.9% (O3-SBBR). Ozonation could convert organic nitrogen and organic substances with strong toxicity and difficult biological utilization into small molecular substances with low toxicity and biodegradability. The abundance of functional genes (nirS, nirK, and nor) in the O3-SBBR combined process was significantly higher than that in the single SBBR, which further confirmed that ozonation could improve the nitrogen removal performance of SBBR. The operation cost of the combined process was 0.74-1.07 yuan·m-3, with good technical economy. This study provided a basis for the application of the O3-SBBR combined process in the advanced treatment of biochemical effluent in printing and dyeing industrial parks.

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The combined denitrifying phosphorus removal (DPR) and Anammox process is expected to achieve advanced nutrient removal with low carbon consumption. However, exchanging ammonia/nitrate between them is one limitation. This study investigated the feasibility of conducting DPR in a biofilm reactor to solve that problem. After 46-day anaerobic/aerobic operation, high phosphorus removal efficiency (PRE, 83.15 %) was obtained in the activated sludge (AS) and biofilm co-existed system, in which the AS performed better. Phosphate-accumulating organisms might quickly adapt to the anoxic introduced nitrate, but the following aerobic stage ensured a low effluent orthophosphate (<1.03 mg/L). Because of waste sludge discharging and AS transforming to biofilm, the suspended solids dropped below 60 mg/L on Day 100, resulting in PRE decline (17.17 %) and effluent orthophosphate rise (4.23 mg/L). Metagenomes analysis revealed that Pseudomonas and Thiothrix had genes for denitrification and encoding Pit phosphate transporter, and Candidatus_Competibacter was necessary for biofilm formation.

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Due to the inhibition of nitrification from aniline toxicity, exogenous N-acyl-homoserine lactones (AHLs) addition was attempted to enhance nitrogen removal in this work. Two sequencing batch biofilm reactors (SBBRs): S1 (the control) and S2 (C6-HSL and 3-oxo-C8-HSL dosing) were used to treat aniline wastewater. The NH4+-N and TN removal rates of S2 were 42.50% and 26.99% higher than S1 in the aerobic phase, respectively. It revealed the nitrogen removal performance of S2 much better than S1. High-throughput sequencing results indicated that many nitrifiers and denitrifiers of S2, such as Nitrosomonas and Thauera, transferred from sludge to biofilm significantly and built closer relationships each other. Overall, main nitrogen removal was contributed by biofilm rather than sludge with the regulation of AHLs. A mild and collaborative environment of biofilms for microorganisms enhanced nitrogen removal. The work provided a new idea for reconciling the contradiction between nitrification and denitrification in aniline wastewater treatment.

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Layered double hydroxides (LDHs) were used as carriers for the microbial consortium in sequencing biofilm batch reactor (SBBR) without inoculation to promote the removal of phosphate. The adsorption capacity of [Zn-Al]-LDH was significantly better than that of [Mg-Al]-LDH. The pollutants removal performance and behavior of microorganisms in LDH-SBBRs were also investigated. LDH-SBBRs showed improved removal efficiencies of COD, phosphate and TP with a low C/N ratio. Microscopic images show that biofilm formed rapidly in LDH-SBBRs. SEM-EDS detected abundant carbon and phosphorus, implying that biomass and phosphorus accumulate on LDH carriers. The microbial compositions of the three SBBRs indicate that the LDHs carriers improved the biodiversity of biofilm in the bioreactors. Synergistic effects of adsorption and biodegradation between well-structured LDHs and microorganisms led to an improved phosphate removal performance of LDH-SBBR. The results also demonstrate that [Zn-Al]-LDH carrier is the best for improving SBBR phosphate removal.

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Generally, the purification performance of bioreactors could be influenced by temperature variation via shaping different microbial communities. However, the underlying mechanisms remain largely unknown. Here, the variation trends of microbial communities in three sequencing batch biofilm reactors (SBBRs) under four different temperatures (15, 20, 25, 30 °C) were compared. It was found that temperature increment led to an obvious enhancement in nutrient removal which was mainly occurred in the aerobic section. Meanwhile, distinct differences in dominant microbial communities or autotrophic nitrifiers were also observed. The performance of the SBBR reactors was closely associated with nitrifier communities since the treated wastewater was characterized by a severe lack of carbon sources (mean effluent COD ≤ 14.4 mg/L). Spearman correlation unraveled that: most of the differentiated microbes as well as the dominant potential functions were strongly associated with nutrient removal, indicating the temperature-induced difference in microbial community well explained the distinction in purification performance.

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Landfill leachate contains many pollutants that have a negative effect on the environment when improperly discharged. Thus the treatment of landfill leachate is a crucial issue, especially in the bigger cities in developing countries. In this study, landfill leachate is treated using a continuous flow sequencing biofilm batch reactor (CF-SBBR) with different biocarriers (non-carrier (NC), kaldness K1 (K1), mutag biochip 30™ (MB), and sponge polyurethane (SP)). The results show that the best COD, TOC, and NH4+-N removal efficiencies were 79.6 ± 0.8%, 78.1 ± 1.9% and 77.5 ± 3.9% in the MB biocarriers tank with an aeration/mixing ratio of 1.3, a cycle time of 9 h and an organic loading rate (OLR) of 1.74 kgCOD/m3.d. The TN removal efficiencies was decreased when there was an increase in the biocarrier's surface area (NC > K1 > MB > SP). At the highest it was 46.1 ± 6.4%, where the aeration/mixing ratio was 1.3, the cycle time was 9 h, and the OLR was 1.52 kgCOD/m3.d. The higher the surface area of the biocarriers, the greater the anti-shock organic loading capacity of the biocarriers due to the formation of biofilm layers. The microbial communities in the CF-SBBR tanks were abundant with common phylum bacteria as in a conventional activated sludge system. Anammox candidatus bacteria was found to total 0.5%. This study concluded that CF-SBBR is an efficient method to treat landfill leachate.

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This study investigates the effectiveness of SBBR with low-cost textile fibers-based bio-carrier namely polypropylene fibers for the treatment of real sewage. The influent loading rates of COD, TN, and TP were averaged at 0.2780, 0.0170, and 0.0077 kg/m3.d, respectively. The removal efficiencies of BOD, COD, TN, and TP recorded in SBBR were 98%, 93%, 82%, and 44%, respectively at an aeration time of 4 h. The TN and TP removal achieved in SBBR were 2.05 and 2.75 times, respectively higher than SBR. The COD removal efficiency was more than 90% under all SRT conditions (10, 14, 18, 22, and 26 d) in SBBR, and the highest efficiency of 93% was obtained at an SRT of 22 days. As the SRT increased, the nitrogen and phosphorus removal decreased, because the denitrification rate and phosphorus release and uptake rate decreased at longer SRT. Simultaneous nitrification and denitrification (SND) efficiency was 85% in SBBR and 44% in SBR, indicating the co-existence of aerobic nitrifiers and anoxic denitrifiers in the biofilm reactor. In SBBR, the nitrogen mass balance showed 74% of nitrogen removed by denitrification, 9% was removed through sludge wasting process, and 13% was removed in effluent at an SRT of 22 days and DO concentration of 3 mg/L. The t-test results suggest that the performance of SBBR was better than SBR in nitrogen and phosphorus removal at a 95% confidence interval.

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A sequencing batch biofilm reactor under intermittent micro-aerobic or anaerobic conditions was investigated to remove pyridine at various concentrations from synthetic wastewater. The results showed that over 98% of pyridine (influent concentration ≤200 mg L-1) was degraded under intermittent micro-aerobic condition, while about 21% of pyridine was removed under anaerobic condition. Additionally, at least 60% of nitrogen located in the pyridine ring was transformed to ammonium. At the same time, the sulphate reduction was obviously inhibited under intermittent micro-aerobic conditions. The microscopic observation showed that abundant microorganisms were attached on the surface or inside of porous biocarriers under intermittent micro-aerobic conditions after a short-term period of operation. High-throughput sequencing analysis demonstrated that Azotobacter, Rhodobacteraceae and Tolumonas were the dominant species in the intermittent micro-aerobic system. The kinetic study at steady period showed that pyridine degradation was fitted well with the pseudo-first-order model (R2 > 0.96). The two possible intermediate products were identified and the possible biodegradation pathway of pyridine was proposed under micro-aerobic condition.

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Marine recirculating aquaculture system (MRAS) is an effective technology that provides sustainable farming of food fish globally. However, dissolved organics material (chemical oxygen demand, COD) and especially ammonia are produced from uneaten feed and metabolic wastes of fish. To purify the MRAS water, this study adopted a sequencing biofilm batch reactor (SBBR) and comparatively investigated the performances of four different carriers on ammonia and COD removal. Results indicated that the NH4+-N removal rates were 0.045 ± 0.05, 0.065 ± 0.008, 0.089 ± 0.005, and 0.093 ± 0.003 kg/(m3·d), and the COD removal rates were 0.019 ± 0.010, 0.213 ± 0.010, 0.255 ± 0.015, and 0.322 ± 0.010 kg/(m3·d) in the SBBRs packed with porous plastic, bamboo ring, maifan stone, and ceramsite carriers, respectively. Among the four carriers, ceramsite exhibited the best performance for both NH4+-N (80%) and COD (33%) removal after the SBBR reached the steady-state operation conditions. For all carriers studied, the NH4+-N removal kinetics could be well simulated by the first-order model, and the NH4+-N and COD removal rates were logarithmically correlated with the carrier's specific surface area. Due to its high ammonia removal, stable performance and easy operation, the ceramsite-packed SBBR is feasible for MRAS water treatment.

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A sequencing biofilm batch reactor (SBBR) running continuously in an anaerobic/aerobic/anoxic (An/O/A) mode was adopted to study the characteristics of nitrification and denitrification process and nitrous oxide (N2O) production under high saline and alkaline conditions. Different carbon and nitrogen ratios (C/N) were also investigated. An influent C/N ratio of 5, 2, and 0 (control), achieved the following results:TN removal efficiency was (98.17±0.42)%, (65.78±2.47)%, and (44.08±0.27)%, respectively; total N2O production was (32.07±2.03) mg·L-1, (21.81±0.85) mg·L-1, and (17.32±0.95) mg·L-1, respectively; and the N2O conversion rate (i. e., the ratio of total N2O production to total nitrogen removal) was (29.75±0.93)%, (30.04±2.17)%, and (41.69±0.80)%, respectively. The nitrification process proceeded normally during the nitrite stage, and nitrite-oxidizing bacteria (NOB) were strongly inhibited under the high saline and alkaline conditions. Due to the inhibition of N2O reductase under these conditions, N2O production was higher during the heterotrophic denitrification process than during the ammonia oxidation process. With an increase in the carbon to nitrogen ratio, more carbon sources were available for denitrification meaning that the total nitrogen removal rate and N2O production were both increased. As the ratio of carbon to nitrogen was increased, the N2O conversion rate decreased, which may have been caused by electron competition among the nitrogen oxide reductases during the denitrification process; the higher the ratio of carbon to nitrogen, the weaker the electron competition. High-throughput sequencing indicated that ammonium-oxidizing bacteria (AOB) were enriched and NOB were almost entirely absent in the SBBR. Thauera, Azoarcus, and Gemmobacter were the dominant heterotrophic denitrifying bacteria identified in the system.

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In this study, controlled C/N effects on fast start-up and stable performance of partial nitrification process at 15℃ in a Sequenced Batch Biofilm Reactor (SBBR) were investigated. The results showed that partial nitrification successfully fast initiated when C/N was 1.5 but failed when C/N was 0/3 during 60 cycles. Fluorescence in situ hybridization and confocal laser scanning microscope (FISH-CLSM) results showed that ammonia oxidizing bacteria (AOB) was found as the dominant bacteria population when C/N was 1.5. When C/N were 0/3, there were almost no existence of AOB and nitrite oxidative bacteria (NOB). Partial nitrification could be stably achieved without carbon source. However, the addition of an appropriate amount of carbon can effectively improve the nitrification performance, and it is better for the stable operation of partial nitrification. In this experiment, partial nitrification was successfully initiated at high dissolved oxygen (DO) (about 9 mg·L-1) conditions. The average DO was maintained at about 6.5 mg·L-1 during the stable operation, which successfully decoupled partial nitrification from low DO concentration. Excessive residual ammonium concentration in the reactors effectively repressed the growth of NOB and guaranteed the stable operation of partial nitrification. At 15℃, full nitritation was more suitable for sidestream wastewater, while mainstream wastewater was more suitable for partial nitritation.

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Oleochemicals industry effluence mainly contains a high chemical oxygen demand (COD) in a range of 6000-20,000 ppm. An effective biological wastewater treatment process must be carried out before wastewater is discharged into the environment. In this study, a submerged bed biofilm reactor (SBBR) was adapted to the biological oleochemical wastewater treatment plant observed in the present study. The effect of wastewater flow rate (100-300 mL/min), Cosmoball® percentage in the SBBR system (25-75%), and percentage of activated sludge (0-50%) were investigated in terms of COD reduction. The Box-Behnken design was used for response surface methodology (RSM) and to create a set of 18 experimental runs, which was needed for optimising the biological oleochemical wastewater treatment. A quadratic polynomial model with estimated coefficients was developed to describe COD reduction patterns. The analysis of variance (ANOVA) shows that the wastewater flow rate was the most effective factor in reducing COD, followed by activated sludge percentage and Cosmoball® carrier percentage. Under the optimum conditions (i.e., a wastewater flow rate of 103.25 mL/min a Cosmoball® carrier percentage of 71.94%, and an activated sludge percentage of 40.50%) a COD reduction of 98% was achieved. Thus, under optimum conditions, as suggested by the BBD, SBBR systems can be used as a viable means of biological wastewater treatment in the oleochemicals industry.

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To enhance the startup and efficient simultaneous nitrification and denitrification for sewage treatment, sequencing batch biofilm reactors (SBBRs) partially coupled with rice husk were established and operated under various intermittent micro-aeration cycles (IMCs) and COD/N ratios under oxygen-limiting intermittent aeration conditions. Experimental results showed that the increase of IMCs with non-aeration/micro-aeration mode of (8 h/4 h)1 to (2 h/1 h)4 in a 12 h-cycle accelerated the startup performance and improved NH4+-N and COD removal. NH4+-N, TN and COD removal efficiencies were 98.7 ± 0.9, 89.2 ± 5.2 and 82.9 ± 6.7% at COD/N ratio of 7.6 with the highest IMCs in SBBR, respectively. Higher TN removal efficiencies of 87.2 ± 4.0 and 58.1 ± 3.5% were also achieved at lower COD/N ratio of 5.6 and 2.8, respectively. In SBBRs with various IMCs, facultative denitrifier like genus Acinetobacter and solid-phase denitrifier belonging to Comamonadaceae family were enriched. However, aerobic denitrifiers with function of heterotrophic nitrification like Paracoccus were favored to enrich under higher IMCs condition, and more anoxic denitrifiers like sulfur-based autotrophic denitrifier Thiothrix and heterotrophic denitrifiers like Pseudomonas and Methyloversatilis were observed at lower IMCs condition. Autotrophic nitrifier (Nitrosomonas and Nitrosipra) and heterotrophic nitrifiers both contributed to the efficient nitrification.

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Milbemycins, a group of 16-membered macrolide antibiotics, are used widely as insecticides and anthelmintics. Previously, a limited understanding of the transcriptional regulation of milbemycin biosynthesis has hampered efforts to enhance antibiotic production by engineering of regulatory genes. Here, a novel ArpA/AfsA-type system, SbbR/SbbA (SBI_08928/SBI_08929), has been identified to be involved in regulating milbemycin biosynthesis in the industrial strain S. bingchenggensis BC04. Inactivation of sbbR in BC04 resulted in markedly decreased production of milbemycin, while deletion of sbbA enhanced milbemycin production. Electrophoresis mobility shift assays (EMSAs) and DNase I footprinting studies showed that SbbR has a specific DNA-binding activity for the promoters of milR (the cluster-situated activator gene for milbemycin production) and the bidirectionally organized genes sbbR and sbbA. Transcriptional analysis suggested that SbbR directly activates the transcription of milR, while represses its own transcription and that of sbbA. Moreover, 11 novel targets of SbbR were additionally found, including seven regulatory genes located in secondary metabolite biosynthetic gene clusters (e.g., sbi_08420, sbi_08432, sbi_09158, sbi_00827, sbi_01376, sbi_09325, and sig24sbh ) and four well-known global regulatory genes (e.g., glnRsbh , wblAsbh , atrAsbh , and mtrA/Bsbh ). These data suggest that SbbR is not only a direct activator of milbemycin production, but also a pleiotropic regulator that controls the expression of other cluster-situated regulatory genes and global regulatory genes. Overall, this study reveals the upper-layer regulatory system that controls milbemycin biosynthesis, which will not only expand our understanding of the complex regulation in milbemycin biosynthesis, but also provide a basis for an approach to improve milbemycin production via genetic manipulation of SbbR/SbbA system.

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The effects of tourmaline on nitrogen removal performance and biofilm structures were comparatively investigated in two identical laboratory-scale sequencing batch biofilm reactors (SBBRs) (denoted SBBR1 and SBBR2) at different nitrogen loading rates (NLRs) varying from (0.24±0.01) to (1.26±0.02) g N/(L·day). SBBR1 was operated in parallel with SBBR2, but SBBR1 was filled with polyurethane foam loaded tourmaline (TPU) carriers and another (SBBR2) filled with polyurethane foam (PU) carriers. Results obtained from this study showed that the excellent and stable performance of SBBR1 was obtained. Ammonia nitrogen removal and total nitrogen removal were higher in SBBR1 than that in SBBR2 with increase of NLR. At an NLR of (0.24±0.01) g N/(L·day), the majority of the spherical and elliptical bacteria were surrounded by the extracellular polymeric substance (EPS) and bacillus or filamentous bacteria in two SBBRs biofilms. When NLR increased to (1.26±0.02) g N/(L·day), the clusters were more obvious in the SBBR1 biofilm than that in the SBBR2 biofilm. Bacteria in SBBR1 were inclined to synthesis more EPS, and the formed EPS could protect the bacteria from free ammonia (FA) under extreme condition NLR (1.26±0.02) g N/(L·day). The results of polymerase chain reaction-denaturing gradient gel electrophoresis analysis showed that the microbial community similarity in SBBR2 decreased more obviously than that in SBBR1 with the increase of NLR, which the microbial community in SBBR1 was relatively stable.

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The effects of florfenicol (FF) on the performance, microbial activity and microbial community of a sequencing batch biofilm reactor (SBBR) were evaluated in treating mariculture wastewater. The chemical oxygen demand (COD) and nitrogen removal were inhibited at high FF concentrations. The specific oxygen utilization rate (SOUR), specific ammonium oxidation rate (SAOR), specific nitrite oxidation rate (SNOR) and specific nitrate reduction rate (SNRR) were decreased with an increase in the FF concentration from 0 to 35 mg/L. The chemical compositions of loosely bound extracellular polymeric substances (LB-EPS) and tightly bound EPS (TB-EPS) could be affected with an increase in the FF concentration. The high-throughput sequencing indicated some obvious variations in the microbial community at different FF concentrations. The relative abundance of Nitrosomonas and Nitrospira showed a decreasing tendency with an increase in the FF concentration, suggesting that FF could affect the nitrification process of SBBR. Some genera capable of reducing nitrate to nitrogen gas could be inhibited by the addition of FF in the influent, such as Azospirillum and Hyphomicrobium.

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The biofilm system is beneficial for Anammox process designed to treat landfill leachate. In this study, the composition of extracellular polymeric substances (EPS) and the microbial community in an Anammox biofilm system were analyzed to determine the functions driving the biofilm's ability to treat landfill leachate. The results demonstrated that increasing influent carbon oxygen demand (COD) could stimulate EPS production. EPS helped enrich Anammox bacteria and supplied them with nutrients and enzymes, facilitating effective nitrogen removal (approximately 95%). The variation in Anammox bacteria was similar to the variation in EPS composition. In the tested Anammox Sequencing Biofilm Batch Reactor (SBBR) system, Candidatus Kuenenia was dominant among known Anammox genus, because of its high substrate affinity and because it adapts better to landfill leachate. The relative abundance of Candidatus Kuenenia in the biofilm rose from 3.26% to 12.38%, illustrating the protection and enrichment offered by the biofilm in carrying out Anammox.

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