Quantification of biofilms in a sub-surface flow wetland and their role in nutrient removal.

Subsurface flow wetlands contain gravel or sand substrates through which the wastewater flows vertically or horizontally. The aims of this study were, firstly, to quantify biofilm development associated with different size gravel in sections of a subsurface flow wetland with and without plants, and secondly, to conduct laboratory experiments to examine the role of biofilms in nutrient removal. Techniques to quantify biofilm included: bacterial cell counts, EPS and total protein extraction. Based on comparative gravel sample volume, only EPS was greater on the smaller 5 mm gravel particles. There was no significant difference between biofilm growth in sections with and without plants. Two vertical flow laboratory-scale reactors, one containing fresh wetland gravel, the other containing autoclaved gravel, were constructed to determine nutrient transformations. The autoclaved gravel in the "sterile" reactor rapidly became colonised with biofilm. Both reactors were dosed with two types of influent. Initially the influent contained 7.25 mg/L NO3-N and 0.3 mg/L NH4-N; the biofilm reactor removed most of the ammonium and nitrite but nitrate concentrations were only reduced by 20%. In the "sterile" reactor there was negligible removal of ammonium and nitrite indicating little nitrification, however nitrate was reduced by 72%, possibly due to assimilatory nitrate reduction associated with new biofilm development. When the influent contained 3 mg/L NO3-N and 16 mg/L NH4-N almost 100% removal and transformation of NH4-N occurred in both reactors providing an effluent high in NO3-N. Organic P was reduced but inorganic soluble P increased possibly due to mineralisation.     

不同氮源对黄花鸢尾净化富营养化水体的影响

利用水生植物床系统研究了不同氮源(硝酸盐、亚硝酸盐、铵盐)对黄花鸢尾(Iris pseudoacorus)去除水体氮磷营养盐效率的影响,同时对植物的生长量、水体中叶绿素a含量、黄花鸢尾对氮磷的吸收利用以及氮循环细菌的分布和氧化亚氮的通量进行了综合研究。结果表明:黄花鸢尾对硝酸盐氮具有优先选择性,而对氨氮的去除效果较差。从植物总氮、总磷吸收量来看,3种氮源中硝酸盐氮>亚硝酸盐氮>氨氮;从氮循环菌分布和N2O释放量来看,硝酸盐氮>氨氮>亚硝酸盐氮。一定范围内,植物对营养盐的吸收随营养盐浓度增加而增加,但水体中营养盐浓度过高则会抑制植物的生长,浓度为80 mg/L的硝酸盐氮,亚硝酸盐氮和氨氮都对黄花鸢尾生长有抑制作用,尤其是高浓度氨氮溶液中,植物的湿重明显减少,因此,黄花鸢尾更适宜治理硝酸盐污染的水体。     

The presence of ammonium facilitates nitrite reduction under PHB driven simultaneous nitrification and denitrification.

For economic and efficient nitrogen removal from wastewater treatment plants via simultaneous nitrification and denitrification the nitrification process should stop at the level of nitrite such that nitrite rather than nitrate becomes the substrate for denitrification. This study aims to contribute to the understanding of the conditions that are necessary to improve nitrite reduction over nitrite oxidation. Laboratory sequencing batch reactors (SBRs) were operated with synthetic wastewater containing acetate as COD and ammonium as the nitrogen source. Computer controlled operation of the reactors allowed reproducible simultaneous nitrification and denitrification (SND). The oxygen supply was kept precisely at a low level of 0.5 mgL(-1) and bacterial PHB was the only electron donor available for denitrification. During SND little nitrite or nitrate accumulated (< 20% total N), indicating that the reducing processes were almost as fast as the production of nitrite and nitrate from nitrification. Nitrite spiking tests were performed to investigate the fate of nitrite under different oxidation (0.1-1.5 mgL(-1) of dissolved oxygen) and reduction conditions. High levels of reducing power were provided by allowing the cells to build up to 2.5 mM of PHB. Nitrite added was preferentially oxidised to nitrate rather than reduced even when dissolved oxygen was low and reducing power (PHB) was excessively high. However, the presence of ammonium enabled significant reduction of nitrite under low oxygen conditions. This is consistent with previous observations in SBR where aerobic nitrite and nitrate reduction occurred only as long as ammonium was present. As soon as ammonium was depleted, the rate of denitrification decreased significantly. The significance of the observed strongly stimulating effect of ammonium on nitrite reduction under SND conditions is discussed and potential consequences for SBR operation are suggested.     

Microbial sulphate reduction during anaerobic digestion: EGSB process performance and potential for nitrite suppression of SRB activity.

The present study investigated mesophilic anaerobic treatment of sulphate-containing wastewater in EGSB reactors and assessed the inclusion of nitrite in the reactor influent as a method for control of biological sulphate reduction. Two EGSB reactors, R1 and R2, were operated for a period of 581 days at varying volumetric loading rates, COD/SO4(2-) ratios and influent nitrite concentrations (R2 only). COD removal efficiencies of > 93% were achieved in both reactors at influent sulphate concentrations of up to 3,000 mg l(-1). A two-fold increase in the influent sulphate concentration, giving an influent COD/SO4(2-) ratio of 2, resulted in a reduction in reactor COD removal efficiency to 84% and 89%, in R1 and R2, respectively. Despite inclusion of nitrite in the R2 influent at concentrations up to 500 mg NO2-N l(-1), sulphate reduction proceeded similarly in R2 and R1, suggesting the ineffectiveness of nitrite as a potential inhibitor of SRB     

Characterization and kinetics of sulfide-oxidizing autotrophic denitrification in batch reactors containing suspended and immobilized cells

Sulfide-oxidizing autotrophic denitrification is an advantageous alternative over heterotrophic denitrification, and may have potential for nitrogen removal of low-strength wastewaters, such as anaerobically pre-treated domestic sewage. This study evaluated the fundamentals and kinetics of this process in batch reactors containing suspended and immobilized cells. Batch tests were performed for different NOx-/S2- ratios and using nitrate and nitrite as electron acceptors. Autotrophic denitrification was observed for both electron acceptors, and NOx-/S2- ratios defined whether sulfide oxidation was complete or not. Kinetic parameter values obtained for nitrate were higher than for nitrite as electron acceptor. Zero-order models were better adjusted to profiles obtained for suspended cell reactors, whereas first-order models were more adequate for immobilized cell reactors. However, in the latter, mass transfer physical phenomena had a significant effect on kinetics based on biochemical reactions. Results showed that sulfide-oxidizing autotrophic denitrification can be successfully established for low-strength wastewaters and have potential for nitrogen removal from anaerobically pre-treated domestic sewage.     

Effect of denitrification type on pH profiles in the sequencing batch reactor process.

Laboratory batch experiments were conducted to investigate pH profiles during partial and complete denitrification with sufficient organic carbon source. Five stirred tank-type glass vessels, with a 7 L working volume for each, were used as SBR reactors that were all operated in denitrification mode. Five levels of initial proportion of nitrogen substances, i.e. nitrate and nitrite, were used in five reactors, respectively. Results showed that, at given temperature and mixed liquor suspended solids (MLSS), partial denitrification could attain a higher pH value than complete denitrification at the end of denitrification with the same initial NOx- concentration. The larger proportion the nitrite took in initial NOx- concentration, the higher pH peak would be obtained on pH profiles during denitrification despite the same total alkalinity produced. It was found that different types of alkalinity were produced during biological denitrification with different nitrogen substances. Partial denitrification could more carbonate alkalinity produce than complete denitrification. Furthermore, some characteristic points were identified on pH profiles which could indicate the disappearance of not only nitrate, but also nitrite in system. When computers are used to detect these features, they can provide rapid, real-time information, regarding the biological state of the system.     

Low temperature biological phosphorus removal and partial nitrification in a pilot sequencing batch reactor system

Partial nitrification and biological phosphorus removal appear to hold promise of a cost-effective and sustainable biological nutrient removal process. Pilot sequencing batch reactors (SBRs) were operated under anaerobic/aerobic configuration for 8 months. It was found that biological phosphorus removal can be achieved in an SBR system, along with the partial nitrification process. Sufficient volatile fatty acids supply was the key for enhanced biological phosphorus removal. This experiment demonstrated that partial nitrification can be achieved even at low temperature with high dissolved oxygen (>3 mg/L) concentration. Shorter solid retention time (SRT) for nitrite oxidizing bacteria (NOB) than for ammonia oxidizing bacteria due to the nitrite substrate limitation at the beginning of the aeration cycle was the reason that caused NOB wash-out. Controlling SRT should be the strategy for an SBR operated in cold climate to achieve partial nitrification. It was also found that the aerobic phosphorus accumulating organisms' P-uptake was more sensitive to nitrite inhibition than the process of anaerobic P-release.     

Nitrogen removal of high strength wastewater via nitritation/denitritation using a sequencing batch reactor.

The sequencing batch reactor (SBR) process concept was applied to achieve efficient ammonium removal via nitrite under both laboratory and pilot-scale conditions. Both sets of experimental results show that without pH control or carbon addition the nitritation process consistently converted approximately 50% of the ammonium from biosolids dewatering liquids to nitrite with hydraulic retention times (HRT) as short as 10 h. The results from the pilot-scale study also indicate that the selective oxidation of ammonium to nitrite is a reliable process as the accumulation of nitrate was never an issue during a 330-day trial. The SBR process concept was extended to achieve complete nitrogen removal through nitritation and denitritation in the laboratory scale. The experimental results indicate that a total reduction of 96-98% of the ammonium nitrogen from biosolids dewatering liquids (influent concentration typically 1,200 g m(-3)) was achieved with a short HRT of 1.1 d and a removal rate of 1.05 kgNm(-3)d(-1). This process concept was tested at pilot scale where the nitritation process could be started up without temperature control in a short period of time. Nitrogen removal rates up to 1.2 kgNm(-3)d(-1) at an HRT of 0.88 d have been obtained. COD to nitrogen ratios required in the pilot plant were consistently in the range 1.6-1.9 kgCOD kg(-1)N removed.     

Leachate pre-treatment strategies before recirculation in landfill bioreactors.

Nitrified leachate recirculation represents a promising strategy for a more sustainable landfill management. Our objective was to determine the reactions involved in nitrate reduction in municipal solid waste batch biodegradation tests. Anaerobic digestion of waste in the three control reactors showed a good reproducibility. In two test reactors, nitrate was added at various moments of the waste degradation process. We observed that: (1) H2S concentration controlled the nitrate reduction pathway: above a certain threshold of H2S, dissimilatory nitrate reduction to ammonium (DNRA) replaced denitrification. (2) N2O/N2 ratio varied with the organic carbon concentration: the lower the easily biodegradable carbon concentration, the higher the N2O/N2 ratio. (3) N2 was consumed after denitrification. The possibility of a nitrogen fixation reaction in the presence of NH4 is discussed. Nitrified leachate recirculation during acidogenesis should be avoided because of higher H2S production which could induce DNRA.     

Anaerobic ammonium oxidation of ammonium-rich waste streams in fixed-bed reactors.

The feasibility of anaerobic ammonium oxidation (Anammox) in fixed-bed reactors was evaluated on laboratory and pilot scales. Using synthetic wastewater, the specific nitrogen removal rate was increased from 0.05-0.1 kgNm(-3)(reactor)d(-1) to 0.35-0.38 kgNm(-3)(reactor)d(-1) within a year (T= 22-27 degrees C) in all applications. However, the anammox activity was seriously and repeatedly inhibited at prolonged high nitrite concentrations (e.g. six days at 30-50 gNO2-Nm(-3)) and recovery was always a lengthy process. But even at a moderate nitrite concentration (11+/-10 gNO2-Nm(-3)), the observed specific growth rate was only 0.018 d(-1) at 26.4+/-0.8 degrees C, which corresponds to approximately 0.025 d(-1) at 30 degrees C (doubling time: 28 days). In a second experimental period for another 250 days, one of the laboratory reactors was fed with partially nitrified sludge liquors from a domestic wastewater treatment plant (WWTP). In this case, the specific elimination rate was as high as 3.5 kgNm(-3)(reactor)d(-1) at 26-27 degrees C. Independently of the feed, the average nitrogen elimination rate lay between 80-85% in all applications. An appropriate hydraulic design is essential to prevent clogging and local nitrite inhibition in fixed-bed reactors.     

Stabilization of source-separated urine by biological nitrification process: treatment performance and nitrite accumulation

A laboratory study on nitrification of high-strength source-separated urine was conducted by means of sequencing batch reactors (SBR) and membrane bioreactors (MBR). The highest influent ammonia concentration for SBR and MBR reached more than 2,400 and 1,000 mg N/L, while the maximum pH was about 9 and 8.9, respectively. The ammonia oxidizing efficiency in both SBRs and MBRs was around 50%, which was restrained mainly by the deficiency of alkalinity in bulks. Meanwhile, the nitrite accumulation did also dominate in these two systems, and the major factor to inhibit the nitrite oxidization was thought to be the high free ammonia and free nitrous acid content in bulks. Hence, an ammonia nitrite solution was achieved with concentration ratio of 1:1; after that ammonia oxidation was restrained owing to the deficiency of alkalinity in urine. The temperature and influent ammonia content have no great influence on the nitrification process in both kinds of bioreactors. The nitrification can be progressed under a solids retention time (SRT) longer than 30 d; however, termination of ammonia oxidization was observed as the SRT fell below 20 d. The nitrifier biomass showed an excellent settleability, such that the suspended solids (SS) in effluent was of a low average, about 60 mg/L. This study on the stabilization of human urine will be useful to understand the process of urine separation from source.     

Partial nitrification of wastewater from an aminoplastic resin producing factory.

Nitrification via nitrite was studied in two aerobic reactors treating wastewater from an aminoplastic resin producing factory at HRT varying between 1.37-1.89 and 2.45-3.63 days. Both eactors were fed with concentrations of 366, 450, 1099 and 1899 mg N-NH4+/L. In general in the reactor operated at a lower HRT, the nitritation percentage decreased from 87.2 to 21.6%, while the nitratation percentage remained always lower than 2.5% (except in the last period) when the ammonium concentration was increased. This behaviour could be due to the inhibition of the ammonium and nitrite oxidation produced by high free ammonia concentrations up to 179.3 mg N-NH3/L. In the reactor operated at a higher HRT, the nitritation percentage decreased and the nitratation percentage increased from 88.6 to 39.6% and from 0.65 to 35.7%, respectively, due to an increase of the dissolved oxygen concentration from 0.76 to 1.02 mg O2/L. However, when ammonium was fed at a concentration of 1898.7 mg N-NH4+/L, the nitritation increased and the nitratation decreased, probably as a result of the accumulation of free ammonia up to 2.04 mg N-NH3/L, meaning that nitrite oxidizers were inhibited. Nitrite build-up was observed after each modification of ammonium concentration in the feed.     

Development of a new type of anaerobic digestion process equipped with the function of nitrogen removal.

In order to develop a new type of anaerobic digestion process equipped with a nitrogen removal function, denitrification of nitrate nitrogen (NO3-N) in anaerobic acidogenesis of organic fraction of municipal waste (OFMSW) was investigated by two semi-continuous reactors. Reactor 1 and Reactor 2 were fed by 3% and 7% of solids concentration of synthetic garbage, respectively. Generation of nitrogen gas (N2) and ammonium nitrogen (NH4-N) was simultaneously observed in the low load of nitrate (NO3-N) (below 0.68 g NO3-N/L). In Reactor 1, ammonium nitrogen generation decreased as the addition of nitrate increased. Finally, the increase of the addition of nitrate resulted in the increase of acetic acid production.     

Effect of inorganic carbon on nitrite accumulation in an aerobic granule reactor.

Pilot scale experiments were performed to evaluate the potential of nitrite type nitrification process with an airlift reactor and granular biomass. Initially, oxygen limitation was used as the main control parameter for accumulating nitrite in the effluent. After 30 d operation, the maximum nitrite conversion rate reached 2.5 kgNO2-N m(-3) d(-1), average diameter of the granule was 0.7 mm. Nitrite type reaction continued over 100 d, but nitrate formation increased after 150 d of operation. Once nitrate formation increased, oxygen limitation could not eliminate nitrite oxidising bacteria from granule. To overcome nitrate formation, laboratory scale batch experiments were conducted and it revealed a high concentration of inorganic carbon which had a significant effect on nitrite accumulation. Following this new concept, inorganic carbon was fed to the pilot scale reactor by changing pH adjustment reagent from NaOH to Na2CO3 and nitrite accumulation was recovered successfully without changing DO concentration. These results show that a high concentration of inorganic carbon is one of the control parameters for accumulating nitrite in biofilm nitrification system.     

Difficulties in maintaining long-term partial nitritation of ammonium-rich sludge digester liquids in a moving-bed biofilm reactor (MBBR).

Nitrogen can be eliminated effectively from sludge digester effluents by anaerobic ammonium oxidation (anammox), but 55-60% of the ammonium must first be oxidized to nitrite. Although a continuous flow stirred tank reactor (CSTR) with suspended biomass could be used, its hydraulic dilution rate is limited to 0.8-1 d(-1) (30 degrees C). Higher specific nitrite production rates can be achieved by sludge retention, as shown here for a moving-bed biofilm reactor (MBBR) with Kaldnes carriers on laboratory and pilot scales. The maximum nitrite production rate amounted to 2.7 gNO2-Nm(-2)d(-1) (3 gO2m(-3)d(-1), 30.5 degrees C), thus doubling the dilution rate compared to CSTR operation with suspended biomass for a supernatant with 700 gNH4-Nm(-3). Whenever the available alkalinity was fully consumed, an optimal amount of nitrite was produced. However, a significant amount of nitrate was produced after 11 months of operation, making the effluent unsuitable for anaerobic ammonium oxidation. Because the sludge retention time (SRT) is relatively long in biofilm systems, slow growth of nitrite oxidizers occurs. None of the selection criteria applied - a high ammonium loading rate, high free ammonia or low oxygen concentration - led to selective suppression of nitrite oxidation. A CSTR or SBR with suspended biomass is consequently recommended for full-scale operation.     

Treatment of nitrogen and phosphorus in highly concentrated effluent in SBR and SBBR processes.

Various sludge treatment processes produced supernatant with high ammonia concentration from 500 to 2,000 mgN/L and generally high phosphate concentration. Conversion of ammonia into nitrite via partial nitrification has proven to be an economic way, reducing oxygen and external COD requirements during the nitrification/denitrification process. Two processes with biomass retention are studied simultaneously: the sequencing batch reactor (SBR) and the sequencing batch biofilm reactor (SBBR). At a temperature of 30 degrees C, the inhibition of nitrite-oxidizing bacteria due to high ammonia concentration has been studied in order to obtain a stable nitrite accumulation. This work has confirmed the effect of pH and dissolved oxygen on nitrite accumulation performance. During a two month starting period, both processes led to nitrite accumulation without nitrate production when pH was maintained above 7.5. From a 500 mgN/L effluent, the performance of the SBR, and the SBBR, reached respectively about 0.95gN-NO2-/gN-NH4+, and 0.4gN-NO2-/gN-NH4+. The SBBR appears to be more stable facing disturbances in dissolved oxygen conditions. Finally, the maximal phosphate removal rates obtained in the SBR reached 90%, and 70% in the SBBR, depending on ammonium accumulation in the reactor. Ammonium phosphate precipitation is likely to occur, as was suggested by crystals observation in the reactor.     

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.     

影响反硝化除磷与厌氧氨氧化耦合的因素

氧对厌氧氨氧化菌有毒,但在颗粒污泥和生物膜中的厌氧氨氧化菌对氧有较高的耐受能力,并且聚磷菌能消耗影响氧氨氧化菌生长的氧。厌氧氨氧化菌的生长无需有机物的参与,聚磷菌释磷需要吸收有机物,少量有机物的加入对厌氧氨氧化菌的活性影响不大。亚硝酸盐是厌氧氨氧化菌氧化氨的电子受体,较高浓度的亚硝酸盐对反硝化聚磷有抑制作用,但合适浓度的亚硝酸盐(该浓度可以通过驯化来提高)可以作为反硝化聚磷菌吸磷的电子受体。厌氧氨氧化过程中有硝酸盐生成,反硝化聚磷菌能利用这部分硝酸盐。另外,两类菌都适宜于中温略偏碱性的环境。因此,通过创造同时对厌氧氨氧化菌和反硝化聚磷菌有利的微生态环境,发挥两者在脱氮除磷方面的协同耦合作用,达到高度脱氮除磷,是极有前景的废水厌氧(缺氧)处理研究方向。     

Factors affecting the activity of anammox bacteria during start up in the continuous culture reactor.

Factors affecting cultivation of extremely slow-growing bacteria (anaerobic ammonium oxidiser, doubling time 11 days) were investigated by using upflow anaerobic sludge blanket (UASB) reactors which can maintain high solid retention time. The effects of concentrations of DO, free ammonia (FA), and nitrite on activation of anammox activity were tested during the start-up period. The reactor was inoculated with granular sludge collected from a full-scale UASB reactor used for treating brewery wastewater, and sludge from a piggery wastewater treatment plant and rotating biological contactor treating sewage. Results of continuous operation showed that concentrations of DO, free ammonia (FA) and nitrite in the reactors played a key role in stimulating the anammox activity during start-up period. It is crucial to keep DO below 0.2 ppm, FA below 2 mg/L and nitrite nitrogen below 35 mg/L to cultivate anammox cells in the continuous bioreactor. When the levels of DO, FA and nitrite in the influent were controlled at less than the inhibition levels, the anammox activity increased gradually in the anaerobic condition. Addition of hydrogen sulphide into the reactor enhanced anammox activity in the continuous culture. Through the SEM, TEM and FISH analysis, anammox bacteria were detected in the granular sludge after 3 months of continuous operation.     

Partial nitrification of sludge reject water by means of aerobic granulation

Granular sludge formation was performed in a laboratory scale Sequencing Batch Reactor (SBR) fed with supernatant of anaerobic digestion of sewage sludge. This effluent was concentrated progressively in order to enhance biomass capacity without inhibiting it. During the first part of the study, ammonium nitrogen was converted to nitrate, so conventional nitrification took place. When a nitrogen load of 0.8 g N L(-1) d(-1) was treated, the effluent concentration of nitrite started to increase while the nitrate concentration decreased until it disappeared. So, partial nitrification was achieved. At the end of this study, a nitrogen load of 1.1 g N L(-1) d(-1) was treated obtaining an effluent with 50% ammonium and 50% nitrite. The volatile suspended solids concentration in the reactor reached 10 g VSS L(-1). At this point the granule morphology was quite round and no filamentous bacteria was observed. The Feret's diameter was in the range between 1 and 6 mm with an average value of 4.5 mm. Roundness value was all the time higher than 0.7. Granule density increased during the experimental period, obtaining a final value of 7.0 g L(-1).