Diversity of ammonia oxidising bacteria in a vertical flow constructed wetland.

Vertical flow constructed wetlands (VFCWs) with intermittent loading are very suitable for nitrification. Ammonia oxidising bacteria (AOB) are the limiting step of nitration. Therefore the AOB community of a full-scale VFCW, receiving municipal wastewater, was investigated within this study. The diversity of the functional gene encoding the alpha-subunit of the ammonia monooxygenase (amoA), present only in AOB, was assessed by denaturing gradient gel electrophoresis (DGGE). Only very few amoA sequence types dominated the wetland filter substrate; nevertheless a stable nitrification performance could be observed. During the cold season the nitrification was slightly reduced, but it has been shown that the same AOB could be identified. No spatial AOB pattern could be observed within the filter body of the VFCW. The most prominent bands were excised from DGGE gels and sequenced. Sequence analyses revealed two dominant AOB lineages: Nitrosomonas europaea/"Nitrosococcus mobilis" and Nitrosospira. Species of the Nitrosomonas lineage are commonly found in conventional wastewater treatment plants (WWTPs). In contrast, members of the Nitrosospira lineage are rarely present in WWTPs. Our observations indicate that the AOB community in this VFCW is similar to that found in horizontal flow constructed wetlands, but differs from common WWTPs regarding the presence of Nitrosospira.     

Performance of an in-situ rotating biological contactor in a recirculating aquaculture system

The start-up and activation of a nitrifying rotating biological contactor (RBC) and its performance inside a culture tank of rainbow trout were studied. First, in a lab-scale operation, the system was fed with a synthetic medium containing a high ammonia concentration (567 mg NH(4)(+)-N L(-1)) and operated at a high hydraulic retention time (HRT) (6.5 days) to minimize the wash-out of the biomass and promote the biofilm formation. Then, both inlet ammonia concentration and HRT were decreased in order to obtain operational conditions similar to those of the culture tank. During this period, the RBC was able to treat an ammonia loading rate (ALR) of 0.64 g N-NH(4)(+) L(-1) d(-1) with a removal efficiency within 70-100%. Pilot-scale experiments were carried out in culture tanks of rainbow trout. The operation of a recirculating system with the RBC unit was compared with a recirculating system without biological treatment and with a flow-through system. The use of this in-situ nitrifying unit allowed working at a recirculation ratio of 90% without negative effects on either growth or the condition factor of fishes. Up to 70% of ammonia generated was removed and a removal rate of 1.41 g NH(4)(+)-N m(-2) d(-1) was reached.     

Nitrogen removal and ammonia-oxidising bacteria in a vertical flow constructed wetland treating inorganic wastewater

Nitrogen removal performance and the ammonia-oxidising bacterial (AOB) community were assessed in the batch loaded 1.3 ha saturated surface vertical flow wetland at CSBP Ltd, a fertiliser and chemical manufacturer located in Kwinana, Western Australia. From September 2008 to October 2009 water quality was monitored and sediment samples collected for bacterial analyses. During the period of study the wetland received an average inflow of 1,109 m3/day with NH3-N = 40 mg/L and NO3-N = 23 mg/L. Effluent NH3-N and NO3-N were on average 31 and 25 mg/L, respectively. The overall NH3-N removal rate for the period was 1.2 g/m2/day indicating the nitrifying capacity of the wetland. The structure of the AOB community was analysed using group specific primers for the ammonia monooxygenase gene (amoA) by terminal restriction fragment length polymorphism and by clone libraries to identify key members. The majority of sequences obtained were most similar to Nitrosomonas sp. while Nitrosospira sp. was less frequent. Another two vertical flow wetlands, 0.8 ha each, were commissioned at CSBP in July 2009, since then the wetland in this study has received nitrified effluent from these two new cells.     

Comparison of nitrification performance and microbial community between submerged membrane bioreactor and conventional activated sludge system.

A submerged membrane bioreactor (SMBR) and a conventional activated sludge system (CAS) were compared in parallel over a period of more than 260 days on treating synthetic ammonia-bearing inorganic wastewater without sludge purge under decreased hydraulic retention times (HRTs). Conversion of NH4(+)-N to NO3(-)-N was achieved with an efficiency of over 98% at an HRT > or = 10 h in the SMBR, while similar performance was obtained at an HRT > or = 20 h in the CAS. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rDNA was used to monitor variations of community structures in the two systems. With the prolongation of operation, the number of DGGE bands in the SMBR gradually increased from the initial 11 bands to the final 22 bands, whereas that in the CAS varied in a range between 13 and 183 Sequence analysis indicates that Nitrosomonas sp. and Nitrospira sp. were the dominating nitrification species responsible for ammonia and nitrite oxidation, respectively. Heterotrophic bacteria like Pseudomonas sp. and Flavobacteria sp. existed in both of the systems although only inorganic wastewater was fed. Substantive accumulation of extracellular polymeric substances (EPS) in the SMBR was confirmed by scanning electron microscopy and EPS analysis.     

Use of real-time PCR to examine the relationship between ammonia oxidizing bacterial populations and nitrogen removal efficiency in a small decentralized treatment system 'Johkasou'.

The aim of this study was to examine the relationship between ammonia oxidizing bacterial populations and biological nitrogen removal in a small on-site domestic wastewater treatment system "Johkasou". The population dynamics of ammonia oxidizing bacteria (AOB) in six full-scale advanced Johkasous was surveyed using real-time PCR assay over a period of one year. These Johkasous were selected to compare the AOB populations in different treatment performance. When the effluent NH4-N concentration was higher than 2 mg L(-1), it was difficult to meet the effluent standard of advanced Johkasous (T-N 10 mg L(-1)). In contrast, the nitrogen removal efficiency was hardly affected by nitrite oxidation and denitrification in these systems. In other words, ammonia oxidation was a rate-limiting step. Furthermore, we focused on the relationship between NH4-N loading per AOB cell and nitrogen removal. Real time PCR monitoring results demonstrated that it is important to regulate NH4-N loading per AOB cell below 210 pg cell(-1) day(-1) to meet the effluent standard of advanced Johkasou. It is considered that NH4-N loading per AOB cell is a useful parameter for determining suitable nitrogen loading and small decentralized system design.     

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.     

Investigating the spatial-temporal variation of nitrogen cycling in an urban river in the North China Plain

Urban rivers are essential in retaining nutrients, but little is known about nitrogen cycling in these rivers in semiarid areas. We measured chemical and isotopic compositions of ammonium (NH4(+)-N) and nitrate (NO3(-)-N) to investigate spatial-temporal variation of nitrogen cycling in the Fuhe River in the North China Plain. Nitrogen pollution in the river was mainly induced by extra NH4(+)-N inputs which come from the discharges of urban sewage and effluents of wastewater treatment plants in upstream. NH4(+)-N obtained from decomposing organic matter of sediments can diffuse into the overlying water. Intense nitrification then occurs at the terrestrial-aquatic interface. Due to less vegetation in spring and autumn, loss of NH4(+)-N is mainly caused by nitrification. In contrast, significant NH4(+)-N is absorbed by plants in summer. NO3(-)-N generated from nitrification can be denitrified during the study period. The highest NO3(-)-N loss (about 86.3%) was observed in summer. The contribution of NO3(-)-N loss due to denitrification is 44.6%. The remaining 55.4% is due to plant uptake. The results suggested that nitrogen cycling in the river is related to temperature and dry-wet cycles. And vegetation restoration along the river could benefit the incremental improvements to the aquatic ecosystem.     

Formation and characteristics of nitritation granules cultivated in sequencing batch reactor by stepwise increase of N/C ratio

The cultivation of nitritation granules in sequencing batch reactor (SBR) by seeding conventional floccular activated sludge was investigated using ethanol-based synthetic wastewater. Reducing settling time offers selection pressure for aerobic granulation, and stepwise increase of influent N/C ratio can help to selectively enrich ammonia oxidizing bacteria (AOB) in aerobic granules. The spherical shaped granules were observed with the mean diameter of 1.25 mm, average settling velocity of 1.9 cm s(-1) and the sludge volume index (SVI) of 18.5-31.4 ml g(-1). After 25 days of operation, the nitrogen loading rate reached 0.0455 kg NH(4)(+)-N (kg MLSS·d)(-1), which was 4.55 times higher than that of the start-up period. The mature granules showed high nitrification ability. Ammonia removal efficiency was above 95% and nitrite accumulation ratio was in the range of 80-95%. The nitrifying bacteria were quantified by fluorescence in situ hybridization analysis, which indicated that AOB was 14.9 ± 0.5% of the total bacteria and nitrite oxidizing bacteria (NOB) was 0.89 ± 0.1% of the total bacteria. Therefore, AOB was the dominant nitrifying bacteria. It was concluded that the associated inhibition of free ammonia at the start of each cycle and free nitrous acid during the later phase of aeration may be the key factors to start up and maintain the stable nitritation.     

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.     

Pilot-scale studies on biological treatment of hypersaline wastewater at low temperature.

In order to investigate the feasibility of biological treatment of hypersaline wastewater produced from toilet flushing with seawater at low temperature, pilot-scale studies were established with plug-flow activated sludge process at low temperature (5-9 degrees C) based on bench-scale experiments. The critical salinity concentration of 30 g/L, which resulted from the cooperation results of the non-halophilic bacteria and the halophilic bacteria, was drawn in bench-scale experiments. Pilot-scale studies showed that high COD removal efficiency, higher than 80%, was obtained at low temperature when 30 percent seawater was introduced. The salinity improved the settleability of activated sludge, and average sludge value dropped down from 38% to 22.5% after adding seawater. Seawater salinity had a strong negative effect on notronomonas and nitrobacter growth, but much more on the nitrobacter. The nitrification action was mainly accomplished by nitrosomonas. Bench-scale experiments using two SBRs were carried out for further investigation under different conditions of salinities, ammonia loadings and temperatures. Biological nitrogen removal via nitrite pathway from wastewater containing 30 percent seawater was achieved, but the ammonia removal efficiency was strongly related not only to the influent ammonia loading at different salinities but also to temperature. When the ratio of seawater to wastewater was 30 percent, and the ammonia loading was below the critical value of 0.15 kgNH4+-N/(kgMLSS.d), the ammonia removal efficiency via nitrite pathway was above 90%. The critical level of ammonia loading was 0.15, 0.08 and 0.03 kgNH4+-N/(kgMLSS.d) respectively at the different temperature 30 degrees C, 25 degrees C and 20 degrees C when the influent ammonia concentration was 60-80 mg/L and pH was 7.5-8.0.     

Treatment of municipal wastewater in a hybrid process using a new suspended carrier with large surface area.

An activated sludge/biofilm hybrid process treating municipal wastewater was studied in pilot plant trials. A new type of suspended carrier, with large effective surface area, was employed in the process with the aim of enhancing nitrification. The pilot plant was operated for 1.5 years in five different configurations including pre-denitrification in all five and enhanced biological phosphorus removal in the final two. The wastewater temperature ranged between 11 degrees C and 20 degrees C, and the nominal dissolved oxygen (DO) level was 5-6 mg/L. The nitrification rate obtained on the new carrier within the hybrid stage was in the range of 0.9-1.2 g NH4-N/m2/d corresponding to a volumetric rate of 19-23 g NH4-N/m3/h (total nitrification including nitrification in the suspended solids). More than 80% of the total nitrification took place on the carrier (and the remainder in the suspended solids). The nitrification rate was shown to correlate with DO, decreasing when the DO was decreased. The results supported the idea of using the new carrier as a tool to upgrade plants not having nitrification today or improve nitrification in activated sludge processes not reaching necessary discharge levels. The large surface area present for nitrification makes it possible to obtain high nitrification rates within limited volumes. The possibility to keep the total suspended solid content low (< 3 g/L) and avoiding problems with the filament Microthrix parvicella, are other beneficial properties of the hybrid process.     

Relevance of the sludge retention time (SRT) as design criteria for wastewater treatment plants for the removal of endocrine disruptors and pharmaceuticals from wastewater.

Wastewater treatment plants (WWTPs) represent a significant source for the input of micro pollutants as endocrine disruptors (EDs) or pharmaceutically active compounds (PhACs) into the aquatic environment. Treatment efficiency of WWTPs often is reported, taking into account only inflow and effluent concentrations without further specification of the WWTP investigated. In order to allow comparison and evaluation of the removal efficiency of different layouts and concepts in wastewater treatment, additional information like the sludge retention time (SRT) and sludge load (F/M ratio) are necessary. Presented results from different WWTPs show correlation of removal of EDs and PhACs to the SRT. Compared to WWTPs with high F/M ratio implementation of the nitrification process on WWTPs results in a significant increase of the removal efficiency for EDs and PhACs. This paper describes an approach to determine comparable removal rates for different activated sludge systems based on mass balance and SRT.     

中水湿地去氮效果研究

以山东省平阴中水湿地为研究对象,通过对湿地水体、底泥的氨态氮(NH3-N)、硝态氮(NO3--N)沿程变化的分析,研究湿地对中水中氮(N)的去除效果。结果表明:湿地对水体中NH3-N的去除效果较好,而作为水体中N主要存在形式的NO3--N沿水流方向却呈增加趋势,微生物硝化作用强烈。底泥中总氮(TN)沿水流方向也呈增加趋势,累计效应比较明显。研究结果为湿地设计的进一步改进提供理论支持。     

Ammonia, a selective agent for methane production by syntrophic acetate oxidation at mesophilic temperature.

In biogas processes, methane production from acetate proceeds by either aceticlastic methanogenesis or through syntrophic acetate oxidation (SAO). In the present study, the pathway for methane production from acetate was analysed; i) during a gradual increase of the ammonia concentration (final concentration 7 g NH(4)(+) -N/L) in a semi-continuous lab-scale anaerobic digester (4.3 L), operating at mesophilic temperature (37 degrees C) or ii) in diluted enrichment cultures (100 ml) experiencing a gradual increase in ammonia, sodium, potassium and propionic acid. The pathway for methane formation was determined by calculating the (14)CO(2)/(14)CH(4) ratio after incubating samples with (14)C-2-acetate. In the anaerobic digester, as well as in the enrichment cultures, the (14)CO(2)/(14)CH4 ratio clearly increased with increasing ammonium-nitrogen concentration, i.e. as the ammonia concentration increased, a shift from the aceticlastic mechanism to the syntrophic pathway occurred. The shift was very distinct and occurred as the NH(4)(+) -N concentration rose above 3 g/l. No shift in pathway was seen during increasing concentrations of sodium, potassium or propionic acid. The shift to SAO in the biogas digester resulted in a twofold decrease in the specific gas and methane yield.     

Performance of slow rate systems for treatment of domestic wastewater.

The performance of slow rate (SR) systems in terms of treatment efficiency, environmental and health risks, and land sustainability was investigated over a three-year period in a rural community close to Iraklio, Greece. Four plant species (Acacia cyanophylla, Eucalyptus camandulensis, Populus nigra and Arundo donax) were used in order to investigate the role of vegetation in the treatment of wastewater and in biomass production. Wastewater effluent was pre-treated in a septic tank before its application to land. Applied hydraulic loading rates were based on crop water requirements which were determined separately for each plant species. The evaluation of treatment performance was accomplished by measuring COD, TKN, NH3-N, NO3-N, total and reactive P, TC and FC in soil solution samples taken at different depths (15, 30 and 60 cm). SR systems showed great potential for COD, TKN and NH4-N removal which reached 89, 90 and 94%, respectively at a depth of 15 cm. An outstanding removal was also observed for TC and FC which reached 99.99%. The concentration of both P and NO3-N in soil solution increased with the passage of time, but it was lower in winter. Despite the differences in the application rates among the SR systems planted with different plant species, the treatment efficiency was not affected. Moreover, increasing the soil depth from 15 to 60 cm had no effect on the treatment efficiency of the SR systems.     

High rate nitrogen removal by the CANON process at ambient temperature

Completely autotrophic nitrogen removal over nitrite (CANON) is a cost-effective nitrogen removal process. Implementation of the CANON process relies on the cooperation of ammonium-oxidizing and Anammox bacteria, as well as the inhibition of nitrite-oxidizing bacteria. Strict limitations on dissolved oxygen (DO) concentration in the reactor, and the addition of sufficient inorganic carbon in the influent, were adopted as the main operational strategies. The reactor was fed with synthetic inorganic wastewater composed mainly of NH(4)(+)-N, and operated for 106 days. Stable nitrogen removal rates (NRR) of around 1.4 kg N m(-3) d(-1) were obtained at ambient temperature. Morphological characteristics and analysis of bacterial community confirmed the formation of functional outer aerobic and inner anaerobic granular sludge, providing evidence of stable nitrogen removal.     

Feasibility study on the removal of nitrous compounds with a hollow-fiber membrane biofilm reactor.

The objective of this study was to develop an integrated nitrogen treatment system using autotrophic organisms. A treatment system consists of an aerobic hollow-fiber membrane biofilm reactor (HfMBR) and anaerobic HfMBR. In the aerobic HfMBR, a mixture gas of air and O2 was supplied through the fibers for nitrification. Denitrification occurred in the anaerobic HfMBR using H2 as the electron donor. The treatment system was continuously operated for 190 days. NH4-N removal efficiencies ranging from 95% to 97% were achieved at NH4-N concentrations of influent ranging from 50 to 100 mg N/L. When glucose was added to the influent, the simultaneous nitrification and denitrification occurred in the aerobic HfMBR, and nitrogen removal rates were changed according to the COD/NH4-N ratio of influent. In the anaerobic HfMBR, autotrophic denitrification using H2 occurred and the removal rates achieved in this study were 23-58 mg N/m2 d. In this study, the achieved removal efficiency was lower than other study findings; however, the result suggested that this hybrid HfMBR system can be used effectively for nitrogen removal in oligotrophic water.     

Experimental study on nitrification in a submerged aerated biofilter.

The aim of the present work was to evaluate the performance of a semi-pilot scale BAF in order to obtain a highly polished effluent in terms of removal of organic matter, suspended solids and ammonia and to observe the influence of temperature, pH and nitrite accumulation on the nitrification process. The ammonia removal efficiency during summer and winter and the nitrite accumulation in presence of free ammonia were observed. The biomass density was measured at different filter bed heights and the sludge production from the effluent of the backwashing water was evaluated. The results obtained were used to calibrate a mathematical model for the prediction of the ammonia removal profile in the filter bed and of biomass thickness.     

基于生态浮床的乡村微污染河道水质净化研究

为了综合解决乡村河流微污染问题,文章以上夹河退水渠河道微污染水体为对象,以净化能力强且观赏性较好的9种水生植物为试验供试植物,构建了单一植物型生态浮床,研究了植物型生态浮床对水体中氨氮(NH4^+-N)、总磷(TP)、化学需氧量(COD)等指标的去除情况,结果表明:9种植物生态浮床对乡村微污染河道水体中污染物的去除率有一定差异;其中,象草、芦苇、水葫芦对NH4^+-N去除率较高,分别为54.58%、61.04%、67.50%;黄菖蒲、美人蕉、水葫芦对TP去除率较高,分别为83.93%、86.26%、89.17%;黄菖蒲、风车草、水葫芦对COD去除率较高,分别为31.82%、31.83%、33.39%;该研究能够为植物型生态浮床技术在今后乡村微污染河流中的应用提供理论指导。     

异养硝化菌的脱氮研究

采用多种选择性培养基进行了异养硝化菌株的筛选,对其生长特性和培养条件进行了研究.随后,又对异养硝化菌强化活性污泥对城市生活污水的处理效果进行了中试研究.试验证明,异养硝化菌能够提高传统污水生物处理的脱氮效率,并有较好的生物絮凝效果,出水氨氮基本监测不出,总氮去除率达80%以上,大大提高了出水水质.