Post-treatment of a slaughterhouse wastewater: stability of the microbial community of a sequencing batch reactor operated under oxygen limited conditions.

Slaughterhouse wastewater is a complex effluent with an important content of organic nitrogen. After an anaerobic treatment where most of the organic matter is removed, the nitrogen, remains as ammonium and post-treatment of the effluent is necessary. Sequencing batch reactor (SBR) technology has been developed to completely remove nitrogen in one single reactor combining aerobic and anoxic stages. Under oxygen limited conditions only nitrite is produced with concomitant energy saving. The stability and diversity of the microbial community from a nitrifying denitrifying SBR operated under oxygen limited conditions were studied using molecular and respirometric methods. The AOB (ammonia oxidizing bacteria) community was relatively stable Nitrosomonas being the dominant genera although Nitrosospira and Nitrosococcus were detected in low proportions. Nitrite oxidizing bacteria were out competed during the operation under oxygen-limited conditions. After an increase of the DO in the reactor Nitrobacter spp were detected suggesting that they remained in the system. Changes in the AOB and denitrifying communities were observed after the DO increase. Sedimentation problems were detected during operation, this could be related to the predominance of Thauera spp detected by FISH and T-RFLP.     

Changes in ammonia oxidiser population during transition to low pH in a biofilm reactor starting with Nitrosomonas europaea.

Recent experiments in our laboratory using both biofilm and suspended biomass reactors have demonstrated high rate nitrification at low pH with known autotrophic nitrifying bacteria originating from wastewater treatment plants refuting previous assumptions that nitrification is significantly inhibited at low pH. Since much of the earlier microbiological work regarding ammonia oxidising bacteria (AOB) physiology was carried out using Nitrosomonas europaea, this model bacterium's capability for high rate nitrification at low pH in a continuous biofilm reactor was tested. A biofilm reactor filled with sintered glass particles was inoculated with a pure culture of N. europaea. The reactor was first operated to high nitrification rates under conditions favourable to N. europaea (pH > 7; high ammonium concentrations). To eliminate inhibitory concentrations of nitrite at low pH, an enriched culture of Nitrospira (a nitrite oxidising bacterium) was then added. The transition from neutral to acidic conditions was attempted by sharply lowering the nitrification rate and by using a feeding solution containing insufficient buffer for complete nitrification. As opposed to other successful transitions, the pH in the N. europaea/Nitrospira reactor initially dropped only slightly and maintained pH > 6 for over two weeks. The reactor reached pH 4.5 only after four weeks. FISH results showed that while the percent of AOB and Nitrospira to eubacteria remained relatively constant at 51.1 +/- 8.2% and 40.8 +/- 6.4%, respectively, the AOB community changed completely in 60 days from 100% N. europaea to 100% Nitrosomonas oligotropha. Even though N. oligotropha was not intentionally introduced into the reactor, it is apparently much better adapted to conditions of low pH.     

Impact of dissolved oxygen concentration on membrane filtering resistance and soluble organic matter characteristics in membrane bioreactors

This study investigated the impact of dissolved oxygen (DO) concentration on membrane filtering resistance, soluble organic matter (SOM) and extracellular polymeric substance (EPS) characteristics in a membrane bioreactor (MBR). A laboratory-scale MBR was operated under DO limited (0.2 mg L(-1) DO) and fully aerobic (3.7 and 5.4 mg L(-1) DO) conditions. Membrane filtering resistance was determined for the mixed liquor suspended solids (MLSS) and for resuspended microbial biomass after removing SOM. Regardless of the DO concentration, the cake resistance (Rc) was approximately 95 percent of the total resistance (Rt). The membrane cake resistance was found to decrease significantly after removing the SOM. The total resistance caused by the resuspended biomass was 29 percent of that caused by the MLSS under DO limited conditions, while the total resistance caused by resuspended biomass was 41 to 48 percent of that caused by the MLSS under fully aerobic conditions. Under DO limited conditions, SOM in the MLSS contained a larger amount of high molecular weight compounds, leading to higher cake resistance than under fully aerobic conditions. There was significant variation in the molecular weight fractions of the EPS, with no clear relationship with DO concentration. There was also no distinct relationship between membrane filtering resistance and molecular weight fraction of the EPS.     

Influence of extracellular polymeric substances on nitrogen removal in an intermittently-aerated membrane bioreactor.

Influence of EPS on fouling of intermittent aeration MBR reactor (denitrification MBR) was investigated changing intermittent aeration cycle (10 minute-cycle and 120 minute-cycle) in laboratory-scale reactors using synthetic wastewater. EPS were extracted from bacterial cells using cation resin method and molecular weight fractioning of EPS was conducted using gel chromatography. In both of the reactors, nitrogen removal rate was almost 100% after 50th day although DO concentration was not very high during the aerated phase because of accumulation of nitrifying bacteria in the reactors. In the 120 minutes-cycle reactor, trans-membrane pressure increased more rapidly than in the 10 minutes-cycle reactor. The reason might be that EPS of more than 1000 kDa, which are the main fouling substances, are produced more rapidly in the 120 minute-cycle condition. It was also found that three peaks at around 100 kDa, 500 kDa and 2000 kDa are prominent in EPS in intermittent-aeration MBR irrespective of cycle and higher molecular weight EPS are decomposed to smaller molecular weight EPS on membrane surface.     

Monitoring pH and ORP in a SHARON reactor

This paper analyses the valuable information provided by the on-line measurements of pH and oxidation reduction potential (ORP) in a continuous single high ammonia removal over nitrite (SHARON) reactor. A laboratory-scale SHARON reactor equipped with pH, ORP, electric conductivity and dissolved oxygen (DO) probes has been operated for more than one year. Nitrogen removal over nitrite has been achieved by adding methanol at the beginning of anoxic stages. Time evolution of pH and ORP along each cycle allows identifying the decrease in nitritation rate when ammonia is consumed during the aerobic phase and the end of the denitrification process during the anoxic phase. Therefore, monitoring pH and ORP can be used to develop a real-time control system aimed at optimizing the length of both aerobic and anoxic stages. Real-time control of methanol addition can be carried out by using the information provided by these probes: excessive methanol addition in the anoxic stage is clearly detected in the ORP profile of the following aerobic phase, while a deficit of methanol is detected in both pH and ORP profiles of that anoxic phase. Moreover, other valuable information such as the amount of ammonia nitrified, failures in DO measurements, excessive stirring during the anoxic stage and methanol dosage in the aerobic phase was also provided by the pH and ORP profiles.     

High nitrification rate at low pH in a fluidized bed reactor with chalk as the biofilm carrier.

A typical steady state bulk pH of about 5 was established in a nitrifying fluidized bed with chalk as the only buffer agent. In spite of the low pH, high rate nitrification was observed with the nitrification kinetic parameters in the chalk reactor similar to those of biological reactors operating at pH>7. Various methods were used to determine the reasons for high rate nitrification at such low pH including (i) determination of bacterial species, (ii) microsensor measurements in the biofilm, and (iii) comparison of nitrification performance at low pH with a non-chalk fluidized bed reactor. Fluorescence in situ hybridization (FISH) using existing 16S rRNA-targeted oligonucleotide probes showed common nitrifying bacteria in the low pH chalk reactor. The prevalent nitrifying bacteria were identified in the Nitrosomonas oligotropha, Nitrosomonas europeae/eutropha, Nitrosospira and Nitrospira related groups, all well known nitrifiers. Microelectrode measurements showed that the pH in the biofilm was low and similar to that of the bulk pH. Finally, reactor performance using a non-chalk biofilm carrier (sintered glass) with the same bacterial inoculum also showed high rate nitrification below pH 5. The results suggest that inhibition of nitrification at low pH is highly overestimated.     

Model-based optimisation of the biological performance of a sidestream MBR.

A model-based optimisation of the operation in view of the biological performance in terms of nitrogen (N) and phosphorus (P) removal of a pilot-scale side-stream MBR has been performed by means of a two-tier scenario analysis. The methodology uses two different scenario analyses to simulate the effect of three degrees of freedom in the MBR system: (1) DO set-point in the aerobic reactor, (2) sludge residence time and (3) internal recirculation rate. The scenarios are simulated using a calibrated ASM2d MBR model. Effluent quality, in terms of nitrate, ammonia and phosphate, is used to select the best scenario. It proved to be a compromise between nitrogen and phosphorus removal as these are linked. A 42% reduction in ammonium and a 32% reduction in nitrate concentration were achieved. Phosphate removal is partly sacrificed (39% increase) compared to the standard operation.     

Alternate anoxic/aerobic operation for nitrogen removal in a membrane bioreactor for municipal wastewater treatment

A large pilot-scale membrane bioreactor (MBR) with a conventional denitrification/nitrification scheme for municipal wastewater treatment has been run for one year under two different aeration strategies in the oxidation/nitrification compartment. During the first five months air supply was provided according to the dissolved-oxygen set-point and the system run as a conventional predenitrification MBR; then, an intermittent aeration strategy based on effluent ammonia nitrogen was adopted in the aerobic compartment in order to assess the impact on process performances in terms of N and P removal, energy consumption and sludge reduction. The experimental inferences show a significant improvement of the effluent quality as COD and total nitrogen, both due to a better utilization of the denitrification potential which is a function of the available electron donor (biodegradable COD) and electron acceptor (nitric nitrogen); particularly, nitrogen removal increased from 67% to 75%. At the same time, a more effective biological phosphorus removal was observed as a consequence of better selection of denitrifying phosphorus accumulating organisms (dPAO). The longer duration of anoxic phases also reflected in a lower excess sludge production (12% decrease) compared with the standard pre-denitrification operation and in a decrease of energy consumption for oxygen supply (about 50%).     

A combined UASB-MBR with shortcut nitrification-denitrification for energy reduction in wastewater reclamation

Shortcut nitrification has been successfully applied in a laboratory scale nitrification-denitrification process consisting of an up-flow anaerobic sludge blanket (UASB) and an aerobic membrane bioreactor (MBR) in treating synthetic and municipal wastewater to simultaneously remove organic carbon and nitrogen. For the treatment of synthetic wastewater, the combined system exhibited a high TOC removal of 98% with a steady ammonia removal efficiency of about 98% in the MBR and a total nitrogen (TN) removal efficiency of 90%. In treating municipal wastewater, due to its low COD concentration, removal efficiencies of TOC, ammonia and TN were 70%, 98% and 60%, respectively. The biogas production was around 76.4 L/m3 wastewater when treating synthetic wastewater. However, little biogas was produced when treating municipal wastewater which was the result of low organic carbon loading to the UASB. Energy analysis has demonstrated that this novel shortcut nitrification process could consume less energy than a conventional process and have the potential of bio-energy generation via biogas production thus helping to achieve a more favorable energy balance.     

Function of dynamic membrane in self-forming dynamic membrane coupled bioreactor.

The Self-Forming Dynamic Membrane Coupled Bioreactor (SFDMBR), which uses coarse pore-sized material to separate solid and liquid in bioreactors, has some advantages compared with MBR using micro-/ultra-filtration membranes, for example, low module cost and high flux. The cake layer and gel layer formed on the surface and in the pores of the material during filtration played an important role, called self-forming dynamic membrane (DM), which mainly consisted of activated sludge. In this study, the function of DM in pollutant removal was investigated. It was found that DM could remove some organic matter (12.6 mg L(-1) on average) and total nitrogen (3.01 mg L(-1) on average) in the supernatant. Colloids and organic nitrogen were partly removed by DM while DOC, ammonia nitrogen and nitrate nitrogen removal by DM varied from negative to positive, which resulted from the combination of various biological activities, e.g. nitrification, biological utilization and so on. DO concentration in DM decreased with the depth and reached zero at about 1.5-2.5 mm depth. The organic degradation activity and nitrification activity of the biomass suspended in the bioreactor were higher than those of the biomass in the cake layer, which might be caused by the low DO concentration and low organic pollutant content in DM.     

曝气条件对侧流活性污泥水解工艺脱氮性能和微生物的影响

通过改变传统厌氧/缺氧/好氧（A²/O）反应器和侧流活性污泥水解（SSH）反应器的曝气强度和溶解氧（DO）浓度,考察了曝气条件对脱氮性能的影响,并对比研究了微生物群落结构的变化规律。结果表明：相较于高DO阶段,两组反应器在中低DO阶段有更好的脱氮效果。在相同进水条件下,SSH反应器的脱氮性能优于A²/O反应器,且出水满足一级A标准。高通量测序结果表明,中低DO浓度更有利于脱氮微生物的生长。相对于A²/O反应器,SSH反应器中反硝化微生物的相对丰度更高。因此,合理控制曝气条件维持中低DO浓度有利于SSH工艺达到良好的脱氮性能及脱氮微生物的生长。     

Nitrification and denitrification in partially aerated biological aerated filter (BAF) with dual size sand media.

A 104-mm (4-inch) diameter pilot-scale biological aerated filter (BAF) with a media depth of 2.5 m (8.3 feet) was operated with an anaerobic, anoxic and oxic zone at a temperature of 23 degrees C. The medium for the anaerobic and anoxic zones was 10 mm diameter sand while the medium for the oxic zone was 5 mm diameter sand. The influent sCOD and total nitrogen concentrations in the feedwater were approximately 250 mg/L and 35 mg N/L, respectively. sCOD removal at optimum hydraulic retention time (HRT) of 3 h with recirculation rates of 100, 200 and 300% in the column was above 96%. Nitrification was found to be more than 96% for 3 h HRT at 200 and 300% recirculation. Total nitrogen removal was consistent at more than 80% for 4 and 6 h HRT at 300% recirculation. For 3 h HRT and 300% recirculation, total nitrogen removal was approximately 79%. The ammonia loading rates for maximum ammonia removed were 0.15 and 0.19 kg NH3-N/m3-day for 100 and 200% recirculation, respectively. The experimental results demonstrated that the BAF can be operated at an HRT of 3h with 200-300% recirculation rates with more than 96% removal of sCOD and ammonia and at least 75% removal of total nitrogen.     

Laboratory study on factors influencing nitrogen removal in marble chip biofilters incorporating nitritation and anammox

It remains challenging to integrate nitritation and anammox in ecologically engineered treatment systems such as passive biofilters that are packed with natural materials and have low energy inputs. This study explored the factors influencing nitritation-anammox through parallel operation of two laboratory-scale biofilters packed with large and small marble chips respectively. Clean marble chips (mainly CaCO3) had an alkalinity dissolution rate of 130 mg CaCO3/kg marble d when water pH approached 6.5. Marble chips effectively increased water pH and provided sufficient alkalinity to support nitritation-anammox in the biofilters. Ammonium and total nitrogen removal decreased by 47 and 26%, respectively, when nutrients were not amended to influent. An influent nitrite concentration above 8.9 mg N/L could inhibit anammox in thin biofilms of biofilters. Nitritation-anammox was enhanced with a hydraulic retention time of 2 d relative to 7 d, likely due to enhanced air entrainment. Size of marble chips rarely made a significant difference in nitrogen removal, possibly due to sufficient surface area available for bacterial attachment and alkalinity dissolution.     

城市污水脱氮除磷SBR在线控制系统研究

SBR采用进水—厌氧—好氧—缺氧—好氧—沉淀—出水的运行方式处理城市污水。反应器装备有DO、ORP和pH等在线检测传感器。DO、ORP和pH变化的一些特征点可以用来判断和控制SBR中污水脱氮除磷过程的各个步骤。这包括:厌氧时,ORP和pH的转折点对应磷的释放;一次好氧时,DO、ORP的氨肘和pH的氨谷对应硝化结束;缺氧时,ORP的硝酸盐膝和pH的硝酸盐峰对应反硝化结束;二次好氧时,DO、ORP碳肘对应剩余碳的氧化结束,pH的转折点对应聚磷结束。控制系统能进行全自动运行来完成污水的脱氮除磷。     

Nitrogen removal from sludge reject water by a two-stage oxygen-limited autotrophic nitrification denitrification process.

Nitrogen removal from sludge reject water was obtained by oxygen-limited partial nitritation resulting in nitrite accumulation in a first stage, followed by autotrophic denitrification of nitrite with ammonium as electron donor (similar to anaerobic ammonium oxidation) in a second stage. Two membrane-assisted bioreactors (MBRs) were used in series to operate with high sludge ages and subsequent high volumetric loading rates, achieving 1.45 kg N m(-3) day(-1) for the partial nitritation MBR and 1.1 kg N m(-3) day(-1) for the anaerobic ammonium oxidation MBR. Biomass retention in the nitritation stage ensured flexibility towards loading rate and operating temperature. Nitrite oxidisers were out-competed at low oxygen and high free ammonia concentration. Biomass retention in the second MBR prevented wash-out of the slowly growing bacteria. Nitrite and ammonium were converted to dinitrogen gas in a reaction ratio of 1.05, thereby maintaining nitrite limitation to assure process stability. The anoxic consortium catalysing the autotrophic denitrification process consisted of Nitrosomonas-like aerobic ammonium oxidizers and anaerobic ammonium oxidizing bacteria closely related to Kuenenia stuttgartiensis. The overall removal efficiency of the combined process was 82% of the incoming ammonium according to a total nitrogen removal rate of 0.55 kg N m(-3) day(-1), without adding extra carbon source.     

Granular biomass capable of partial nitritation and anammox

A novel and efficient way of removing nitrogen from wastewater poor in biodegradable organic carbon, is the combination of partial nitritation and anoxic ammonium oxidation (anammox), as in the one-stage oxygen-limited autotrophic nitrification/denitrification (OLAND) process. Since anoxic ammonium-oxidizing bacteria grow very slowly, maximum biomass retention in the reactor is required. In this study, a lab-scale sequencing batch reactor (SBR) was used to develop granular, rapidly settling biomass. With SBR cycles of one hour and a minimum biomass settling velocity of 0.7 m/h, OLAND granules were formed in 1.5 months and the nitrogen removal rate increased from 50 to 450 mg N L(-1) d(-1) in 2 months. The granules had a mean diameter of 1.8 mm and their aerobic and anoxic ammonium-oxidizing activities were well equilibrated to perform the OLAND reaction. Fluorescent in-situ hybridization (FISH) demonstrated the presence of both beta-proteobacterial aerobic ammonium oxidizers and planctomycetes (among which anoxic ammonium oxidizers) in the granules. The presented results show the applicability of rapidly settling granular biomass for one-stage partial nitritation and anammox.     

Full-cyclic control strategy of SBR for nitrogen removal in strong wastewater using common sensors.

A full-cyclic automatic control strategy for sequencing batch reactors (SBR) was proposed using only common sensors such as ORP, DO and pH. The main objective was to develop a generally applicable and robust control strategy. To accomplish this, various control schemes found in the literature or suggested by authors were examined at diverse ammonia loads and SCOD/NH4(+)-N ratios. Advantages and constraints of each scheme were discussed and compared. Ammonia load was estimated with DO lag time during the aerobic stage, and then the influent pump was manipulated to meet the desired load at the next anoxic stage. A partial denitrification scheme was chosen for the anoxic stage period control, to save anoxic time and external carbon. For external carbon dosage control, intermittent feeding at each anoxic stage was concluded to be a suitable scheme. The anoxic stage period could be successfully controlled by the combination of pH increase and DO increase. Every suggested control scheme was incorporated into a full-cyclic control strategy and tested at 0.02, 0.035, 0.08 kg NH4(+)-N/m3/sub-cycle. From the results, it is expected to perform unmanned automatic SBR operation with this strategy.     

Simultaneous nitrification and de-nitrification in MBR.

Experiments have been carried out to get an understanding of the effect of DO, C/N ratio and pH on the performance of a bench scale membrane bioreactor (MBR) in simultaneous nitrification and denitrification. It was found that under the conditions of MLSS in the range of 8000-9000 mg/L and temperature of water in the MBR of 24 degrees C, influent COD and NH3-N in the range of 523-700 mg/L and 17.24-24 mg/L respectively, the removals of COD, NH3-N and TN were 98%, 99% and 60%; 96.5%, 0,98% and 75%; 96%, 95% and 92%; 90%,70% and 60% respectively at DO of 6, 3, 1 and 0.5 mg/L. It was also found that the changes in C/N ratio and pH in a certain range have a slight effect on COD removal but have significant influence on the removal of NH3-N and TN. The results showed that only under the conditions that each ecological factor was maintained relatively steadily, simultaneous nitrification and de-nitrification proceeded smoothly. It was found that when C/N ratio was 30, the influent pH 7.2, the temperature of water in MBR 24 degrees C and DO 1 mg/L, as optimum conditions, the removals of COD, NH3-N and TN were 96%, 95% and 92% respectively. In addition, mechanism research on simultaneous nitrification and de-nitrification in MBR has been conducted as well.     

Fluorescence in situ hybridization analysis of nitrifiers in piggery wastewater treatment reactors.

Fluorescence in situ hybridization (FISH) was performed to analyze the nitrifying microbial communities in an activated sludge reactor (ASR) and a fixed biofilm reactor (FBR) for piggery wastewater treatment. Heterotrophic oxidation and nitrification were occurring simultaneously in the ASR and the COD and nitrification efficiencies depend on the loads. In the FBR nitrification efficiency also depends on ammonium load to the reactor and nitrite was accumulated when free ammonia concentration was higher than 0.2 mg NH3-N/L. FISH analysis showed that ammonia-oxidizing bacteria (NSO1225) and denitrifying bacteria (RRP1088) were less abundant than other bacteria (EUB338) in ASR. Further analysis on nitrifying bacteria in the FBR showed that Nitrosomonas species (NSM156) and Nitrospira species (NSR1156) were the dominant ammonia-oxidizing and nitrite-oxidizing bacteria, respectively, in the piggery wastewater nitrification system.     

Control of nutrients after discharge to lakes through wastewater.

Control of nutrients as nitrogen and phosphorus after discharge into lakes is necessary since it is difficult and costly to control within wastewater plants currently in China. This paper studied the cycling of phosphorus and nitrogen with water and sediments from two lakes in China. It is found that oxygen plays a critical role in regulating phosphorus and nitrogen cycling within water and sediments. Three different oxygenation methods including aeration, calcium peroxide and hydrogen peroxide were studied to control phosphorus and nitrogen in overlying water. In anoxic conditions, the P concentration in water increased from an average 14 microg/L to 115.2 microg/L for Xili Lake, and from an average 24 microg/L to 1,000 microg/L for Jinchun Lake. The concentration of ammonia increased under anoxic conditions, while the concentration of nitrate increased under oxic conditions. In anoxic conditions, the nitrate concentration decreased probably through denitrification. Both N and P accumulation processes can be controlled under the three treatments. The phosphorus removal efficiency from the water body was in the order of CaO2 addition > aeration > H2O2 addition, while controlling effectiveness for ammonia was in the order of aeration > CaO2 addition > H2O2 addition.