Simultaneous nitrification–denitrification and microbial community profile in an oxygen-limiting intermittent aeration SBBR with biodegradable carriers

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.     

Aerobic granulation with low strength wastewater at low aeration rate in A/O/A SBR reactor

The formation of aerobic granules with low organic loading synthetic wastewater (150-200 mg L⁻¹ of influent COD, acetate/propionate = 1/3) at low aeration rate (0.6 cm s⁻¹ of superficial gas velocity) had been investigated in the anaerobic/oxic/anoxic SBR. Aerobic granules with smooth surface and compact structure were successfully obtained after 50 days. However, these aerobic granules were unstable when the d(0.9) of granules increased to more than 1 mm. The results suggested that the aerobic granules with small diameter (smaller than 1000 μm) were more favorable for treating the low substrate loading wastewater at the low aeration rate. The cycle test revealed that most of the influent COD was removed at the anaerobic stage. The effluent concentrations of N-NH₄⁺ and P-PO₄³⁻ were lower than 1 mg L⁻¹, and the effluent concentration of nitrate gradually decreased with the granulation. Phosphate accumulating organisms were found to utilize O₂ or NO_x⁻ as electron acceptor for phosphorus removal in the study. Simultaneous nitrogen and phosphorus removal occurred inside the granules.     

Characteristics of granular methanogenic sludge grown on glucose in a UASB reactor

The formation of granules grown on glucose in an upflow anaerobic sludge blanket (UASB) reactor was investigated. Total granular sludge concentration retained in the UASB reactor was 34.5 g MLSS/l (30.0 g MLVSS/l) during 240 d operation on glucose minimum medium with the supplementation of 1.07 g NaHCO₃ per 1 g glucose. This realized a high-rate methanogenic fermentation of glucose of 17.6 g COD/l-reactor-d at 3.4 d⁻¹ of space velocity. The granules formed were relatively small, ranging mainly from 0.4 to 0.5 mm, had a relatively low cell density of 0.0542–0.0560 g MLVSS/ml, and had low specific gravity (0.97–1.19) due to very low ash content (11–13%). Electron microscopic analysis showed that Methanothrix spp. appeared dominant over the granules. The specific metabolic activities of bacterial trophic groups were the highest for H₂ followed by glucose, acetate, and propionate.     

Effect of Mn<Superscript>2+</Superscript> augmentation on reinforcing aerobic sludge granulation in a sequencing batch reactor

Two sequencing batch reactors were synchronously operated to investigate the effect of manganese (II) (Mn²⁺) augmentation on aerobic granulation. Reactor 1 (R1) was added with 10 mg/L Mn²⁺, while there was no Mn²⁺ augmentation in reactor 2 (R2). Results showed that R1 had a faster granulation process than R2 and R1 performed better in chemical oxygen demand (COD) and ammonium nitrogen (NH₄⁺–N) removal efficiencies. Moreover, the mature granules augmented with Mn²⁺ behaved better on their physical characteristics and size distributions, and they also had higher production of extracellular polymeric substances (EPS) content. The result of three-dimensional excitation and emission matrix fluorescence showed that Mn²⁺ had the function of causing organic material diversity (especially proteins diversity) in EPS fraction from granules. Polymerase chain reaction and denaturing gradient gel electrophoresis techniques were employed to analyze the microbial and genetic characteristics in mature granules. The results exhibited that Mn²⁺ augmentation was mainly responsible for the higher microbial diversity of granules from R1 compared with that from R2. Uncultured sludge bacterium A16 (AF234726) and Rhodococcus sp. WTZ-R2 (HM004214) were the major species in R1, while only uncultured sludge bacterium A16 (AF234726) in R2. Moreover, there were eight species of organisms found in both two aerobic granules, and three species were found only in aerobic granules from R1. It could be concluded that Mn²⁺ could enhance the sludge granulation process and have a key effect role on the biological properties during the sludge granulation.     

Enhanced aerobic granulation and nitrogen removal by the addition of zeolite powder in a sequencing batch reactor

The aim of this study was to evaluate the impact of zeolite powders on feasibility of rapid aerobic granulation in the column-type sequencing batch reactors. After 90 days' operation, aerobic granular sludge was formed in both reactors by altering influent chemical oxygen demand/nitrogen (COD/N) ratios. R1 with zeolite powders had better removal capabilities of COD and total nitrogen than R2, which was without zeolite powders. Mixed liquor volatile suspended solid concentrations of the two reactors were 7.36 and 5.45 g/L, while sludge volume index (SVI₃₀) values were 34.9 and 47.9 mg/L, respectively. The mean diameters of aerobic granular sludge in the above two reactors were 2.5 and 1.5 mm, respectively. Both reactors achieved the largest simultaneous nitrification and denitrification (SND) efficiency at an influent COD/N ratio of 8; however, R1 exhibited more excellent SND efficiency than R2. The obtained results could provide a novel technique for rapid aerobic granulation and N removal simultaneously, especially when treating nitrogen-rich industrial wastewater.     

Nitrogen removal via simultaneous partial nitrification,anammox and denitrification (SNAD) process under high DO condition

The simultaneous partial nitrification, anammox and denitrification (SNAD) process for treating domestic wastewater was investigated in a sequencing batch reactor (SBR). The SBR was operated with air flow rate of 500 L h^?1 at 30 °C. Domestic wastewater was used as influent and Kaldnes rings were used as biomass carriers. In the beginning, long aeration condition was implemented to cultivate nitrification biofilm. Afterwards, intermittent aerobic condition was conducted during the cycle operation. The influent organic matter loading rate was improved by reducing the aeration and mixing times. Consequently, when the SNAD biofilm reactor was fed with the organic matter loading rate of 0.77 (kg COD m^?3 d^?1), the bio-bubbles appeared in the reactor and the total inorganic nitrogen (TIN) removal efficiency decreased. After the organic matter loading rate decreased to 0.67 (kg COD m^?3 d^?1), the reactor showed excellent nitrogen removal performance. The TIN removal efficiency varied between 80 and 90 %, and the average TIN removal loading rate was 0.22 (kg TIN m^?3 d^?1). Additionally, the scanning electron microscope (SEM) observation confirmed that the anammox bacteria located in the inner part of the carriers. Finally, the microbial community analysis of 16S rRNA gene cloning revealed that the anammox bacteria on the carriers consisted of three main genuses: Candidatus Brocadia sp., Candidatus Brocadia caroliniensis and Candidatus Brocadia fulgida.     

Investigation on the formation and kinetics of glucose-fed aerobic granular sludge

Aerobic granular sludge was cultivated in a glass sequencing batch reactor (SBR) with glucose synthetic wastewater. The spherical shaped granules were observed on 4th day with the mean diameter of 0.1 mm. With the increase of chemical oxygen demand (COD) concentration of the influent, aerobic granules grew matured, the size of which ranged from 1.2 to 1.9 mm. The aerobic granular sludge could sustain high organic loading rate (about 4.0 g COD L⁻¹ d⁻¹), with good settling ability (settling velocity 36 m/h) and high biomass concentration (MLSS 6.7 ±0.2 g/L). Experimental data indicated that the substrate utilization and biomass growth kinetics followed Monod's kinetics model approximately. The corresponding kinetic coefficients of maximum specific substrate utilization rate (k), half velocity coefficient (K_s), growth yield coefficient (Y) and decay coefficient (K_d) were 13.2 d⁻¹, 275.8 mg/L, 0.183–0.250 mg MLSS/mg COD and 0.023–0.075 d⁻¹, respectively, which made aerobic granules have short setup period, high rate of substrate utilization and little surplus sludge.     

Nitrogen removal from wastewater with a low COD/N ratio at a low oxygen concentration

The goal of the study was to determine the effectiveness of nitrification and denitrification and the kinetics of ammonia removal from a mixture of wastewater and anaerobic sludge digester supernatant in an SBR at limited oxygen concentration. In addition, the COD removal efficiency and sludge production were assessed.In the SBR cycle alternating aerobic and anaerobic phases occurred; in the aeration phase the dissolved oxygen (DO) concentration was below 0.7 mg O₂/L. The low DO concentration did not inhibit ammonia oxidation-nitrification and the efficiency was ca. 96-98%. However, a relatively high COD concentration in the effluent was detected. The values of K_m and V_max, calculated from the Michaelis-Menten equation, were 43 mg N-NH₄/L and 15.64 mg N-NH₄/L h, respectively. Activated sludge production was almost stable (0.62-0.66 g MLVSS/g COD). A high net biomass production resulted from a low specific biomass decay rate of 0.0015 d⁻¹.     

Performance of bench-scale membrane bioreactor under real work conditions using pure oxygen: viscosity and oxygen transfer analysis

Pure oxygen to supply the aerobic condition was used in the performance of a bench-scale submerged membrane bioreactor (MBR). The pilot plant was located in the wastewater treatment plant of the city of Granada (Spain) and the experimental work was divided into two stages (Unsteady state and steady state conditions). Operation parameters (MLSS, MLVSS and dissolved oxygen concentration) and physical characteristics (temperature, conductivity, pH, COD and BOD₅) were daily monitored. The results showed the capacity of the MBR systems to remove organic material under a hydraulic retention time of 18.46 h and sludge retention time of 18.6 days. Therefore, Viscosity of the sludge and αkLa-factor of the aeration, were determinate in the steady stage condition to understand the behavior of the system when pure oxygen has been used to supply the aerobic conditions of the MBR system showed an alpha-factor of 0.238 when the viscosity of the system was 4.04 Cp.     

Characteristics of aerobic granular sludge in a sequencing batch reactor with variable aeration

Aerobic granules can be used for the treatment of industrial or municipal wastewater, but high aeration rate is required for the stable operation of the granular sludge system. Therefore, the aim of this research was to reduce aeration rate greatly to decrease the energy consumption for the technology of aerobic granules. Based on the characteristics of sequencing batch reactor with distinct feast and famine periods, aeration rate was reduced from 1.66 to 0.55 cm s⁻¹ in the famine period after granules were formed. It was found that the settleability of aerobic granules in reactor R1 with reduced aeration was the same as that of aerobic granules in reactor R2 with constant aeration rate of 1.66 cm s⁻¹. However, the outer morphology of aerobic granules gradually changed from round shape to long shape, and minor population showed certain shift after aeration rate was reduced in the famine period. Since good settleability is the most essential feature of aerobic granules, it can be said that reducing aeration rate in famine period did not influence the stable operation of aerobic granular sludge system. Furthermore, the experimental results indicated that aeration rate in feast period was much more important to the stable operation of aerobic granules than that in famine period.     

Characteristics of granular methanogenic sludge grown on phenol synthetic medium and methanogenic fermentation of phenolic wastewater in a UASB reactor

The growth of granules on a phenol synthetic medium and the methanogenic fermentation of industrial phenolic wastewater from a steel factory in an upflow anaerobic sludge blanket (UASB) reactor were investigated. Total granular sludge concentration retained in the UASB reactor was 6.7 g MLSS/l (6.0 g MLVSS/l) during the 10 months' operation on the phenol synthetic medium. This realized a maximum phenol removal rate of 2.2 g/l·d (phenol concentration of influent = 500 mg/l), which corresponded to 5.2 g COD/l·d at space velocity (SV) of 4.4 d⁻¹. The granules formed were of relatively small size ranging from 0.61 to 0.77 mm, and had a relatively low density of 0.013–0.023 g MLVSS/cm³ and low specific gravity (1.11) due to very low ash content (8.7–11.9%). Electron microscopic analysis showed that Methanothrix spp. appeared dominantly on the granule surface as well as within it. The specific metabolic activities of bacterial trophic groups were the highest for H₂ followed by acetate, benzoate, phenol, and propionate. In the case of industrial phenolic wastewater, although phenol efficiency was only 50% at SV of 0.4 d⁻¹, when the wastewater was diluted twofold and the treated wastewater was recycled at SV of 7.3 d⁻¹, the removal efficiencies of phenol and CODcr were restored to 90% (influent=400 mg/l) and 80% (influent=5,000 mg/l), respectively. It was suggested that recycling of the treated wastewater might be improved by partly degrading unknown toxic compounds contained in phenolic wastewater.     

Potential cause of aerobic granular sludge breakdown at high organic loading rates

Aerobic sludge granules are compact, strong microbial aggregates that have excellent settling ability and capability to efficiently treat high-strength and toxic wastewaters. Aerobic granules disintegrate under high organic loading rates (OLR). This study cultivated aerobic granules using acetate as the sole carbon and energy source in three identical sequencing batch reactors operated under OLR of 9–21.3 kg chemical oxygen demand (COD) m⁻³ day⁻¹. The cultivated granules removed 94–96% of fed COD at OLR up to 9–19.5 kg COD m⁻³ day⁻¹, and disintegrated at OLR of 21.3 kg COD m⁻³ day⁻¹. Most tested isolates did not grow in the medium at >3,000 mg COD l⁻¹; additionally, these strains lost capability for auto-aggregation and protein or polysaccharide productivity. This critical COD regime correlates strongly with the OLR range in which granules started disintegrating. Reduced protein quantity secreted by isolates was associated with the noted poor granule integrity under high OLR. This work identified a potential cause of biological nature for aerobic granules breakdown.     

Starvation is not a prerequisite for the formation of aerobic granules

Activated sludge with sludge volume index (SVI)₃₀ of 77 ml g⁻¹ and SVI₃₀ of 433 ml g⁻¹ was inoculated to start up reactors R1 and R2, respectively. In both R1 and R2, cycle time of 1 h and the influent chemical oxygen demand (COD) concentrations of 1,000 mg l⁻¹ were employed. Initial settling time of 2 min resulted in the loss of a substantial amount of biomass as wash-out and high effluent COD concentrations within the first week of operation. This implied that there was no starvation phase in each cycle of R1 and R2 during the first week of operation. However, aerobic granules with a size above 400 μm formed by day 7. Thus, it was concluded that starvation was not a prerequisite for the formation of aerobic granules. When cycle time was 1 h, the instability of aerobic granules was observed. When cycle time was prolonged to 1.5 h and granular sludge of 200 ml was used to start up reactor R3, the reactor R3 reached steady state within 1 week. SVI, size, and the morphology of granular sludge in R3 remained stable during the 47-day operation, which indicated that prolonged starvation time had positive effects on the stability of aerobic granules.     

Sequence optimization in a sequencing batch reactor for biological nutrient removal from domestic wastewater

The purpose of this work was to determine optimum sequence retention times for nutrient removal with low-cost using very short aeration time in an SBR treating domestic wastewater. During the study, four different CYCLEs were evaluated, with the highest removal efficiencies recorded for the CYCLE with fill, anaerobic, aerobic1, anoxic, aerobic2, settle, and decant sequences operated at retention times of 0.5, 2, 2, 1, 0.75, 1, and 0.5 h, respectively. For this CYCLE, the removal efficiencies of chemical oxygen demand (COD), total Kjeldahl nitrogen (TKN), ammonia nitrogen (NH₃–N), total phosphorus (TP), and ortho-phosphate (PO₄–P) were found, on average, to be 91, 78, 85, 87, and 83%, respectively. The optimum sequence retention time was determined via the analysis of variance (ANOVA) using the Matlab software (Mathworks Inc.). The data indicated that the total time of the aerobic sequences was shorter than those of previous studies for similar level of removal efficiencies in all parameters including N and P.     

Biodegradation and kinetics of aerobic granules under high organic loading rates in sequencing batch reactor

Biodegradation, kinetics, and microbial diversity of aerobic granules were investigated under a high range of organic loading rate 6.0 to 12.0 kg chemical oxygen demand (COD) m⁻³ day⁻¹ in a sequencing batch reactor. The selection and enriching of different bacterial species under different organic loading rates had an important effect on the characteristics and performance of the mature aerobic granules and caused the difference on granular biodegradation and kinetic behaviors. Good granular characteristics and performance were presented at steady state under various organic loading rates. Larger and denser aerobic granules were developed and stabilized at relatively higher organic loading rates with decreased bioactivity in terms of specific oxygen utilization rate and specific growth rate (μ _overall) or solid retention time. The decrease of bioactivity was helpful to maintain granule stability under high organic loading rates and improve reactor operation. The corresponding biokinetic coefficients of endogenous decay rate (k _d), observed yield (Y _obs), and theoretical yield (Y) were measured and calculated in this study. As the increase of organic loading rate, a decreased net sludge production (Y _obs) is associated with an increased solid retention time, while k _d and Y changed insignificantly and can be regarded as constants under different organic loading rates.     

Variable aeration in sequencing batch reactor with aerobic granular sludge

This study investigated the effects of reduced aeration in famine period on the performance of sequencing batch reactor (SBR) with aerobic granular sludge. Aerobic granules were first cultivated in two SBRs (R1 and R2) with acetate as sole carbon source. From operation day 27, aeration rate in R1 was reduced from 1.66 to 0.55 cm s(-1) from 110 min to the end of each cycle and further reduced from 30 min to the end of each cycle from day 63. R2 as a control was operated with a constant aeration rate of 1.66 cm s(-1) in the whole cycle during the entire experimental period. Results showed that changing trends of SVI, concentration, average size and VSS/SS of biomass with time in R1 and R2 were similar although different aeration modes were adopted. At steady state, SVI of aerobic granules and biomass concentration maintained at about 40 ml g(-1) and 6 g l(-1), respectively. Average size of granules was about 750 microm in R1 while 550 microm in R2. This is the first study to demonstrate that aerobic granular sludge could be stable at reduced aeration rate in famine period during more than 3-month operation. Such an operation strategy with reduced aeration rate will lead to a significant reduction of energy consumption, which makes the aerobic granular sludge technology more competitive over conventional activated sludge process. Furthermore, the stability of aerobic granular system with variable aeration further indicates that the difference of physiology and kinetics of aerobic granule in feast and famine periods results in the different requirements of oxygen and shear stress for the stability of granules, which will deepen the understanding of mechanism of aerobic granulation in sequencing batch reactor.     

Characteristics of methanogenic granules grown on propionate in an upflow anaerobic sludge blanket (UASB) reactor

Granulation of a propionate-degrading consortia was performed with a mesophilic propionate-acclimatized sludge in an upflow anaerobic sludge blanket (UASB) reactor. The granules formed were relatively small, ranging mainly from 0.3 to 0.6 mm in diameter, but had an excellent sedimentation velocity due to a high specific gravity of 1.355 g/cm³ (ash content, 48.2%). The ash consisted mainly of calcium (30.2%), phosphorus (19.7%), and magnesium (3.95%) forming plate crystals in the granules. The populations of three bacterial trophic groups present in the granules, propionate-degraders, hydrogenotrophic and aceticlastic methanogens were 5.6 × 10⁸, 1.6 × 10¹⁰, and 2 × 10⁹ (in most probable number/g mixed-liquor volatile suspended solids [MLVSS]), respectively, while the specific utilization rates of propionate, hydrogen, and acetate of the granules were 9.4, 850, and 20.9 (mmol/g MLVSS·d), respectively. Electron microscopic analysis showed that Methanothrix spp. appeared dominant over the granules. Total granular sludge concentration retained in the UASB reactor during 178 d of operation was 80.0 g mixed-liquor suspended solids (MLSS)/l-reactor, corresponding to 41.4 g MLVSS/l-reactor, which realized a high-rate methanogenic fermentation of propionate of 85 g chemical oxygen demand (COD)/l-reactor·d.     

Enhanced aerobic granulation with extracellular polymeric substances (EPS)-free pellets

Extracellular polymeric substances (EPSs) were secreted by cells after they agglomerated into a compact aggregate. This study shows that the EPS initially embedded in seed sludge before granulation may sterically slow subsequent microbe–microbe contact, thereby delaying aerobic granulation. Three identical bioreactors were used in this study using glucose as the sole carbon and energy source. Reactor 1 (R1) was seeded with EPS-free pellets and operated in sequencing batch reactor (SBR) mode. Reactor 2 (R2) was seeded with the original sludge flocs and operated in SBR mode. Reactor 3 (R3) was seeded with EPS-free pellets and operated in continuously stirred tank reactor (CSTR) mode. Granulation occurred in R1 earlier than in R2; the granules that formed in R1 were larger and more compact than those in R2 at the same cultivation time. The few mature granules in R3 suggest that aerobic granulation can occur in a CSTR when a reactor is seeded with EPS-free pellets.     

Production of hydrogen from domestic wastewater in a pilot-scale microbial electrolysis cell

Addressing the need to recover energy from the treatment of domestic wastewater, a 120-L microbial electrolysis cell was operated on site in Northern England, using raw domestic wastewater to produce virtually pure hydrogen gas (100?±?6.4 %) for a period of over 3 months. The volumetric loading rate was 0.14 kg of chemical oxygen demand (COD) per cubic metre per day, just below the typical loading rates for activated sludge of 0.2–2 kg?COD?m^?3?day^?1, at an energetic cost of 2.3 kJ/g?COD, which is below the values for activated sludge 2.5–7.2 kJ/g?COD. The reactor produced an equivalent of 0.015 L?H₂?L^?1?day^?1, and recovered around 70 % of the electrical energy input with a coulombic efficiency of 55 %. Although the reactor did not reach the breakeven point of 100 % electrical energy recovery and COD removal was limited, improved hydrogen capture and reactor design could increase the performance levels substantially. Importantly, for the first time, a ‘proof of concept’ has been made, showing that this technology is capable of energy capture as hydrogen gas from low strength domestic wastewaters at ambient temperatures.     

Treatment of old landfill leachate with high ammonium content using aerobic granular sludge

Background

Aerobic granular sludge has become an attractive alternative to the conventional activated sludge due to its high settling velocity, compact structure, and higher tolerance to toxic substances and adverse conditions. Aerobic granular sludge process has been studied intensively in the treatment of municipal and industrial wastewater. However, information on leachate treatment using aerobic granular sludge is very limited.

Methods

This study investigated the treatment performance of old landfill leachate with different levels of ammonium using two aerobic sequencing batch reactors (SBR): an activated sludge SBR (ASBR) and a granular sludge SBR (GSBR). Aerobic granules were successfully developed using old leachate with low ammonium concentration (136 mg L^?1 NH₄ ⁺-N).

Results

The GSBR obtained a stable chemical oxygen demand (COD) removal of 70% after 15 days of operation; while the ASBR required a start-up of at least 30 days and obtained unstable COD removal varying from 38 to 70%. Ammonium concentration was gradually increased in both reactors. Increasing influent ammonium concentration to 225 mg L^?1 N, the GSBR removed 73 ± 8% of COD; while COD removal of the ASBR was 59 ± 9%. The GSBR was also more efficient than the ASBR for nitrogen removal. The granular sludge could adapt to the increasing concentrations of ammonium, achieving 95 ± 7% removal efficiency at a maximum influent concentration of 465 mg L^?1 N. Ammonium removal of 96 ± 5% was obtained by the ASBR when it was fed with a maximum of 217 mg L^?1 NH₄ ⁺-N. However, the ASBR was partially inhibited by free-ammonia and nitrite accumulation rate increased up to 85%. Free-nitrous acid and the low biodegradability of organic carbon were likely the main factors affecting phosphorus removal.

Conclusion

The results from this research suggested that aerobic granular sludge have advantage over activated sludge in leachate treatment.