Development and characteristics of phosphorus-accumulating microbial granules in sequencing batch reactors

Phosphorus (P)-accumulating microbial granules were developed at different substrate P/chemical oxygen demand (COD) ratios in the range of 1/100 to 10/100 by weight in sequencing batch reactors. The soluble COD and PO4-P profiles showed that the granules had typical P-accumulating characteristics, with concomitant uptake of soluble organic carbon and the release of phosphate in the anaerobic stage, followed by rapid phosphate uptake in the aerobic stage. The size of P-accumulating granules exhibited a decreasing trend with the increase in substrate P/COD ratio, while the structure of the granules became more compact and denser as the substrate P/COD ratio increased. The P uptake by granules fell within the range of 1.9% to 9.3% by weight, which is comparable with uptake obtained in conventional enhanced biological phosphorus removal (EBPR) processes. It was further found that low aerobic respirometric activity of granules in terms of specific oxygen utilization rate favors P uptake by granules. The results presented would be useful for the further development of a novel granule-based EBPR technology.     

Modeling and experimental study on the anaerobic/aerobic/anoxic process for simultaneous nitrogen and phosphorus removal: The effect of acetate addition

A mathematical model based on the simulation software AQUASIM was developed to validate an anaerobic/aerobic/anoxic (AOA) process that enables simultaneous nitrogen and phosphorus removal in a single reactor by adding external organic carbon to preclude excess aerobic phosphate uptake by polyphosphate-accumulating organisms (PAOs) and provide phosphate for denitrifying PAOs (DNPAOs). Aerobic batch tests after anaerobic phosphate release with different chemical oxygen demand (COD) concentrations indicated that the effect of COD concentration on the phosphate uptake preclusion could be expressed by a simple formula. The reduction factor reflecting the formula, which retards the aerobic phosphate uptake in the presence of COD, was added to the process rates of aerobic polyphosphate storage and PAOs growth in the model. The improved model, which included the reduction factor, reasonably matched the experimental result regarding aerobic phosphate uptake behavior whereas the model without it did not; thus, the former precisely predicts the AOA process behavior.     

Simultaneous nitrification,denitrification, and phosphorus removal in a lab-scale sequencing batch reactor

Simultaneous nitrification and denitrification (SND) via the nitrite pathway and anaerobic-anoxic-enhanced biological phosphorus removal (EBPR) are two processes that can significantly reduce the energy and COD demand for nitrogen and phosphorus removal. The combination of these two processes has the potential of achieving simultaneous nitrogen and phosphorus removal with a minimal requirement for COD. A lab-scale sequencing batch reactor (SBR) was operated in alternating anaerobic-aerobic mode with a low dissolved oxygen (DO) concentration (0.5 mg/L) during the aerobic period, and was demonstrated to accomplish nitrification, denitrification, and phosphorus removal. Under anaerobic conditions, COD was taken up and converted to polyhydroxyalkanoates (PHAs), accompanied by phosphorus release. In the subsequent aerobic stage, PHA was oxidized and phosphorus was taken up to <0.5 mg/L by the end of the cycle. Ammonia was also oxidized during the aerobic period, but without accumulation of nitrite or nitrate in the system, indicating the occurrence of simultaneous nitrification and denitrification. However, off-gas analysis showed that the final denitrification product was mainly nitrous oxide (N(2)O), not N(2). Further experimental results demonstrated that nitrogen removal was via nitrite, not nitrate. These experiments also showed that denitrifying glycogen-accumulating organisms (DGAOs), rather than denitrifying polyphosphate-accumulating organisms (DPAOs), were responsible for the denitrification activity.     

Simultaneous nitrification, denitrification, and phosphorus removal from nutrient-rich industrial wastewater using granular sludge

The biological removal of nitrogen and phosphorus from nutrient-rich abattoir wastewater using granular sludge has been investigated. A lab-scale sequencing batch reactor, seeded with granular sludge developed using synthetic wastewater, was operated for 13 months under alternating anaerobic and aerobic conditions. It is demonstrated that the granules could be sustained and indeed further developed with the use of abattoir wastewater. The organic, nitrogen, and phosphorus loading rates applied were 2.7 gCOD L(-1) day(-1), 0.43 gN L(-1) day(-1), and 0.06 gP L(-1) day(-1), respectively. The removal efficiency of soluble COD, soluble nitrogen and soluble phosphorus were 85%, 93%, and 89%, respectively. However, the high suspended solids in the effluent limited the overall removal efficiency to 68%, 86%, and 74% for total COD, TN, and TP, respectively. This good nutrient removal was achieved through the process known as simultaneous nitrification, denitrification, and phosphorus removal, likely facilitated by the presence of large anoxic zones in the center of the granules. The removal of nitrogen was likely via nitrite optimizing the use of the limited COD available in the wastewater. Accumulibacter spp. were found to be responsible for most of the denitrification, further reducing the COD requirement for nitrogen and phosphorus removal. Mineral precipitation was evaluated and was not found to significantly contribute to the overall nutrient removal. It is also shown that the minimum HRT in a granular sludge system is not governed by the sludge settleability, as is the case with floccular sludge systems, but likely by the limitations associated with the transfer of substrates in granules.     

Kinetic model of a granular sludge SBR: influences on nutrient removal

A mathematical model was developed that can be used to describe an aerobic granular sludge reactor, fed with a defined influent, capable of simultaneously removing COD, nitrogen and phosphate in one sequencing batch reactor (SBR). The model described the experimental data from this complex system sufficiently. The effect of process parameters on the nutrient removal rates could therefore be reliably evaluated. The influence of oxygen concentration, temperature, granule diameter, sludge loading rate, and cycle configuration were analyzed. Oxygen penetration depth in combination with the position of the autotrophic biomass played a crucial role in the conversion rates of the different components and thus on overall nutrient removal efficiencies. The ratio between aerobic and anoxic volume in the granule strongly determines the N-removal efficiency as it was shown by model simulations with varying oxygen concentration, temperature, and granule size. The optimum granule diameter for maximum N- and P-removal in the standard case operating conditions (DO 2 mg L(-1), 20 degrees C) was found between 1.2 and 1.4 mm and the optimum COD loading rate was 1.9 kg COD m(-3) day(-1). When all ammonia is oxidized, oxygen diffuses to the core of the granule inhibiting the denitrification process. In order to optimize the process, anoxic phases can be implemented in the SBR-cycle configuration, leading to a more efficient overall N-removal. Phosphate removal efficiency mainly depends on the sludge age; if the SRT exceeds 30 days not enough biomass is removed from the system to keep effluent phosphate concentrations low.     

Feasibility of nitrification/denitrification in a sequencing batch biofilm reactor with liquid circulation applied to post-treatment

An investigation was performed on the biological removal of ammonium nitrogen from synthetic wastewater by the simultaneous nitrification/denitrification (SND) process, using a sequencing batch biofilm reactor (SBBR). System behavior was analyzed as to the effects of sludge type used as inoculum (autotrophic/heterotrophic), wastewater feed strategy (batch/fed-batch) and aeration strategy (continuous/intermittent). The presence of an autotrophic aerobic sludge showed to be essential for nitrification startup, despite publications stating the existence of heterotrophic organisms capable of nitrifying organic and inorganic nitrogen compounds at low dissolved oxygen concentrations. As to feed strategy, batch operation (synthetic wastewater containing 100 mg COD/L and 50 mg N-NH(4)(+)/L) followed by fed-batch (synthetic wastewater with 100 mg COD/L) during a whole cycle seemed to be the most adequate, mainly during the denitrification phase. Regarding aeration strategy, an intermittent mode, with dissolved oxygen concentration of 2.0mg/L in the aeration phase, showed the best results. Under these optimal conditions, 97% of influent ammonium nitrogen (80% of total nitrogen) was removed at a rate of 86.5 mg N-NH(4)(+)/Ld. In the treated effluent only 0.2 mg N-NO(2)(-)/L,4.6 mg N-NO(3)(-)/L and 1.0 mg N-NH(4)(+)/L remained, demonstrating the potential viability of this process in post-treatment of wastewaters containing ammonium nitrogen.     

Characterization of aerobic granules by microbial density at different COD loading rates

Aerobic granules were cultivated under temporal alternating aerobic and anoxic conditions without the presence of a carrier material in a sequencing batch reactor (SBR) with a high column height/column diameter ratio. The reactor was operated for 6h per cycle (aerobic: 4.75 h, anoxic: 1.25 h). To determine a new parameter for the definition of aerobic granules, a protocol of 4,6-diamidino-2-phenylindole hydrochloride staining and fluorescence image processing was developed. The d(tm) analysis showed that the increase in the chemical oxygen demand (COD) loading rate promoted no more growth of the aerobic granules. It was inconsistent with the results of the analysis of the sludge volume index (SVI) value but matched well with the results of the COD and nitrogen removal of the SBR and the particle size distribution by LS-PSA. The optimum COD loading rate for aerobic granulation in the SBR was 2.52 kg/m(3)d. When d(tm) was correlated with the biomass concentration and the SVI value during the period of granule formation, d(tm) could be used as a more sensitive and accurate parameter for classifying aerobic granules and optimizing the operational conditions for aerobic granulation processes.     

Organic and nitrogen removal in a two-stage rotating biological contactor treating municipal wastewater

A laboratory scale rotating biological contactor (RBC) predenitrification system incorporating anoxic and aerobic units was evaluated for the treatment of settled high-strength municipal wastewater. The system was operated under four recycle ratios (1, 2, 3 and 4) and loading rates of 38-182 gCOD/m(2)d and 0.22-14 gOxid-N/m(2)d on the anoxic unit and 3.4-18 gCOD/m(2)d and 0.24-1.8 gNH(4)-N/m(2)d on the aerobic. The average removal efficiency in terms of chemical oxygen demand (COD), biochemical oxygen demand (BOD(5)), total suspended solids (TSS) and total nitrogen (Total-N) was 82%, 86%, 63% and 54%; settling of the RBC effluent increased COD and TSS removal to 94% and 97%. An increase in hydraulic loading resulting from higher recirculation, had limited negative effect on organic removal but improved nitrogen removal, and in terms of Total-N removal efficiency increased up to a ratio of 3 and then decreased.     

反硝化聚磷菌及其脱氮除磷机理研究进展

水体富营养化是当前水环境保护工作的重点关注问题,微生物修复富营养化水体具有高效、低耗且不产生二次污染等特点,已经成为富营养化水体生态修复的一种重要方式。近年来,对反硝化聚磷菌的研究及其在污水处理工艺中的应用越来越广泛。不同于传统的反硝化细菌联合聚磷菌去除氮磷工艺,反硝化聚磷菌在交替厌氧、缺氧/好氧条件下能同时进行脱氮除磷而被广泛关注与研究。值得注意的是,近几年报道的部分微生物仅在好氧条件下就可进行同时脱氮除磷,但是其脱氮除磷机理仍未理清。基于此,文中总结了目前发现的反硝化聚磷菌和同时硝化反硝化聚磷微生物的种类及特点,并对其脱氮与除磷的关系及其机理进行了系统性分析,对目前反硝化除磷存在的问题进行了梳理,最后对今后的研究方向进行了展望,以期为完善反硝化聚磷菌的脱氮除磷机理及工艺改进提供参考。     

The investigation of effect of organic carbon sources addition in anaerobic–aerobic (low dissolved oxygen) sequencing batch reactor for nutrients removal from wastewaters

The effect of addition of organic carbon sources (acetic acid and waste activated sludge alkaline fermentation liquid) on anaerobic–aerobic (low dissolved oxygen, 0.15–0.45 mg/L) biological municipal wastewater treatment was investigated. The results showed that carbon source addition affected not only the transformations of polyhydroxyalkanoates (PHA), glycogen, nitrogen and phosphorus, but the net removal of nitrogen and phosphorus. The removal efficiencies of TN and TP were, respectively, 61% and 61% without organic carbon source addition, 81% and 95% with acetic acid addition, and 83% and 97% with waste activated sludge alkaline fermentation liquid addition. It seems that the alkaline fermentation liquid of waste biosolids generated in biological wastewater treatment plant can be used to replace acetic acid as an additional carbon source to improve the anaerobic–aerobic (low dissolved oxygen) municipal wastewater nutrients removal although its use was observed to cause a slight increase of effluent BOD and COD concentrations.     

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.     

Experimental investigation of the external nitrification biological nutrient removal activated sludge (ENBNRAS) system

A systematic lab-scale experimental investigation is reported for the external nitrification (EN) biological nutrient removal (BNR) activated sludge (ENBNRAS) system, which is a combined fixed and suspended medium system. The ENBNRAS system was proposed to intensify the treatment capacity of BNR-activated sludge (BNRAS) systems by addressing two difficulties often encountered in practice: (a) the long sludge age for nitrification requirement; and (b) sludge bulking. In the ENBNRAS system, nitrification is transferred from the aerobic reactor in the suspended medium activated sludge system to a fixed medium nitrification system. Thus, the sludge age of the suspended medium activated sludge system can be reduced from 20 to 25 days to 8 to 10 days, resulting in a decrease in reactor volume per ML wastewater treated of about 30%. Furthermore, the aerobic mass fraction can also be reduced from 50% to 60% to <30% and concommitantly the anoxic mass fraction can be increased from 25% to 35% to >55% (if the anaerobic mass fraction is 15%), and thus complete denitrification in the anoxic reactors becomes possible. Research indicates that both the short sludge age and complete denitrification could ameliorate anoxic aerobic (AA) or low food/microorganism (F/M) ratio filamentous bulking, and hence reduce the surface area of secondary settling tanks or increase the treatment capacity of existing systems. The lab-scale experimental investigations indicate that the ENBNRAS system can obtain: (i) very good chemical oxygen demand (COD) removal, even with an aerobic mass fraction as low as 20%; (ii) high nitrogen removal, even for a wastewater with a high total kjeldahl nitrogen (TKN)/COD ratio, up to 0.14; (iii) adequate settling sludge (diluted sludge volume index [DSVI] <100 mL/g); and (iv) a significant reduction in oxygen demand.     

The development of simultaneous partial nitrification, ANAMMOX and denitrification (SNAD) process in a single reactor for nitrogen removal

The simultaneous partial nitrification, ANAMMOX and denitrification (SNAD) process was validated to potentially remove ammonium and COD from wastewater in a single, oxygen-limited, non-woven rotating biological contactor (NRBC) reactor. An ammonium conversion efficiency of 79%, TN removal efficiency of 70% and COD removal efficiency of 94% were obtained with the nitrogen and COD loading rate of 0.69 kgN/m(3)d and 0.34 kg/m(3)d, respectively. Scanning electron microscopy (SEM) observation and fluorescence in situ hybridizations (FISH) analysis revealed the existence of the dominant groups of bacteria. As a result, the aerobic ammonia-oxidizing bacteria (AOB), with a spot of aerobic heterotrophic bacteria were mainly distributed in the aerobic outer part of the biofilm. However, ANAMMOX bacteria with denitrifying bacteria were present and active in the anaerobic inner part of the SNAD biofilm. These bacteria were found to exist in a dynamic equilibrium to achieve simultaneous nitrogen and COD removal in NRBC system.     

Partial nitrification and nutrient removal in intermittently aerated sequencing batch reactors treating separated digestate liquid after anaerobic digestion of pig manure

The performance of an intermittently aerated sequencing batch reactor (IASBR) technology was investigated in achieving partial nitrification, organic matter removal and nitrogen removal from separated digestate liquid after anaerobic digestion of pig manure. The wastewater had chemical oxygen demand (COD) concentrations of 11,540 ± 860 mg/L, 5-day biochemical oxygen demand (BOD₅) concentrations of 2,900 ± 200 mg/L and total nitrogen (TN) concentrations of 4,041 ± 59 mg/L, with low COD:N ratios (2.9) and BOD₅:COD ratios (0.25). Synthetic wastewater, simulating the separated digestate liquid with similar COD and nitrogen concentrations but BOD₅ of 11,500 ± 100 mg/L, was also treated using the IASBR technology. At a mean organic loading rate of 1.15 kg COD/(m³ d) and a nitrogen loading rate of 0.38 kg N/(m³ d), the COD removal efficiency was 89.8% in the IASBR (IASBR-1) treating digestate liquid and 99% in the IASBR (IASBR-2) treating synthetic wastewater. The IASBR-1 effluent COD was mainly due to inert organic matter and can be further reduced to less than 40 mg/L through coagulation. The partial nitrification efficiency of 71–79% was achieved in the two IASBRs and one cause for the stable long-term partial nitrification was the intermittent aeration strategy. Nitrogen removal efficiencies were 76.5 and 97% in IASBR-1 and IASBR-2, respectively. The high nitrogen removal efficiencies show that the IASBR technology is a promising technology for nitrogen removal from low COD:N ratio wastewaters. The nitrogen balance analysis shows that 59.4 and 74.3% of nitrogen removed was via heterotrophic denitrification in the non-aeration periods in IASBR-1 and IASBR-2, respectively.     

Identifying an interfering factor on chemical oxygen demand (COD) determination in piggery wastewater and eliminating the factor by an indigenous Pseudomonas stutzeri strain

AIMS: This study attempted to demonstrate nitrite interference on chemical oxygen demand (COD) determination in piggery wastewater, and the capability of aerobic denitrification of the SU2 strain which is capable of promoting the efficiency of nitrogen and COD removal from piggery wastewater. METHODS AND RESULTS: This study was performed in a 17-litre reactor with a 30% packing ratio, with a ratio of immobilized SU2 cells to sludge of 100:1. The ratio of aeration to nonaeration was 4 : 1.5. Removal efficiency of COD was 86.8%. Removal efficiency of BOD and SS was higher than 90%, and removal efficiency of NH4+-N and TKN was almost 100%. CONCLUSIONS: NO2- -N interference is significant when its concentration in piggery wastewater exceeds 100 mg l-1. COD in piggery wastewater can be indirectly reduced following nitrite reduction by SU2 strain. SIGNIFICANCE AND IMPACT OF THE STUDY: Utilizing immobilized SU2 cells in coordination with an SBR system simultaneously reduces nitrite and COD concentrations.     

Biomass granulation in an aerobic:anaerobic-enhanced biological phosphorus removal process in a sequencing batch reactor with varying pH

Long-term influences of different steady-state pH conditions on microbial community composition were determined by fluorescence in situ hybridization (FISH) in a laboratory scale reactor configured for enhanced biological phosphorus removal (EBPR). Chemical profiles were consistent with shifts in populations from polyphosphate-accumulating organisms (PAO) to glycogen-accumulating organisms (GAO) when pH fell from pH 7.5 to 7.0 and then to 6.5. While biomass was both dispersed and flocculated at pH 7.5, almost complete granulation occurred gradually after pH was dropped to 7.0, and these granules increased in size as the pH was reduced further to 6.5. Reverting back to pH 7.5 led to granule breakdown and corresponding increases in anaerobic phosphate release. Granules consisted almost entirely of Accumulibacter PAO cells, while putative GAO populations were always present in small numbers. Results suggest that low pH may contribute to granulation under these operational conditions. While chemical profiles suggested the PAO:GAO balance was changing as pH fell, FISH failed to reveal any marked corresponding increase in GAO abundances. Instead, TEM evidence suggested the Accumulibacter PAO phenotype was becoming more like that of a GAO. These data show how metabolically adaptable the Accumulibacter PAO can be under anaerobic:aerobic conditions in being able to cope with marked changes in plant conditions. They suggest that decreases in EBPR capacity may not necessarily reflect shifts in community composition, but in the existing population metabolism.     

Protozoan biomass relation to nutrient and chemical oxygen demand removal in activated sludge mixed liquor

The relationship between biomass concentration to nutrient and chemical oxygen demand (COD) removal in mixed liquor supplemented with sodium acetate was investigated, using three protozoan isolates and three different initial biomass concentrations (10(1), 10(2) and 10(3) cells/mL). The study was carried out in a shaking flask environment at a shaking speed of 100 rpm for 96 h at 25 degrees C. Aliquot samples were taken periodically for the determination of phosphate, nitrate, COD and dissolved oxygen, using standard methods. The results revealed remarkable phosphate removal of 82-95% at biomass concentration of 10(3)cells/mL. A high nitrate removal of over 87% was observed at all initial biomass concentration in mixed liquor. There was an observed COD increase of over 50% in mixed liquor in at the end of 96-h incubation and this was irrespective of initial biomass concentration used for inoculation. The study shows the trend in nutrient and COD removal at different biomass concentrations of the test isolates in mixed liquor.     

Control of COD/N ratio for nutrient removal in a modified membrane bioreactor (MBR) treating high strength wastewater

A modified membrane bioreactor (MBR) system has been developed to evaluate the efficiency of nutrient removal in treating synthetic high strength water. This study examined the effect of influent COD/N ratio on this system. Results showed that above 95.0% removal efficiencies of organic matter were achieved; indicating COD removal was irrespective of COD/N ratio. The average removal efficiencies of total nitrogen (TN) and phosphate (PO(4)(3-)-P) with a COD/N ratio of 9.3 were the highest at 90.6% and 90.5%, respectively. Furthermore, TN removal was primarily based on simultaneous nitrification and denitrification (SND) process occurred in the aerobic zone. Decreased COD/N ratios to 7.0 and 5.3, TN removal efficiencies in steady-states were 69.3% and 71.2%, respectively. Both aerobic SND and conventional biological nitrification/denitrification contributed to nitrogen removal and the latter played dominant effect. PO(4)(3-)-P-release and uptake process ceased in steady-states of COD/N 7.0 and 5.3, which decreased its removal efficiency significantly.     

Effect of COD/N ratio on cultivation of aerobic granular sludge in a pilot-scale sequencing batch reactor

Aerobic granular sludge was successfully cultivated with the effluent of internal circulation reactor in a pilot-scale sequencing batch reactor (SBR). Soy protein wastewater was used as an external carbon source for altering the influent chemical oxygen demand/nitrogen (COD/N) ratios of SBR. Initially, the phenomenon of partial nitrification was observed and depressed by increasing the influent COD/N ratios from 3.32 to 7.24 mg/mg. After 90 days of aerobic granulation, the mixed liquor suspended solids concentration of the reactor increased from 2.80 to 7.02 g/L, while the sludge volumetric index decreased from 105.51 to 42.99 mL/g. The diameters of mature aerobic granules vary in the range of 1.2 to 2.0 mm. The reactor showed excellent removal performances for COD and $ {\text{NH}}_4^{ + }{\text{ - N}} $ after aerobic granulation, and average removal efficiencies were over 93% and 98%, respectively. The result of this study could provide further information on the development of aerobic granule-based system for full-scale applications.