Start-up of the anammox process from the conventional activated sludge in a hybrid bioreactor

The anaerobic ammonium oxidation (anammox) process was successfully started up from conventional activated sludge using a hybrid bioreactor within 2 months. The average removal efficiencies of ammonia and nitrite were both over 80%, and the maximum total nitrogen removal rate of 1.85 kg N/(m³.day) was obtained on day 362 with the initial sludge concentration of 0.7 g mixed liquor suspended solids (MLSS)/L. Scanning electron microscope (SEM) observation of the granular sludge in the hybrid reactor clearly showed a high degree of compactness and cell sphericity, and the cell size was quite uniform. Transmission electron microscope photos showed that cells were round or oval, the cellular diameter was 0.6--1.0 μupm, and the percentage of the anammoxosome compartment was 51%--85% of the whole cell volume. Fluorescence in situ hybridization analysis (FISH) indicated that anammox bacteria became the dominant population in the community (accounting for more than 51% of total bacteria on day 250). Seven planctomycete 16S rRNA gene sequences were present in the 16S rRNA gene clone library generated from the biomass and affiliated to Candidatus Kuenenia stuttgartiensis and Candidatus Brocadia sp., a new anammox species. In addition, the average effluent suspended solid (MLSS) concentrations of outlets I (above the non-woven carrier) and II (below the non-woven carrier) were 0.0009 and 0.0035 g/L, respectively. This showed that the non-woven carrier could catch the biomass effectively, which increased biomass and improved the nitrogen removal rate in the reactor.     

Influence of seasonal temperature change on autotrophic nitrogen removal for mature landfill leachate treatment with high-ammonia by partial nitrification-Anammox process

In this study, a denitrification (DN)–partial nitritation (PN)–anaerobic ammonia oxidation (Anammox) system for the efficient nitrogen removal of mature landfill leachate was built with a zone-partitioning self-reflux biological reactor as the core device, and the effects of changes in seasonal temperature on the nitrogen removal in non-temperature-control environment were explored. The results showed that as the seasonal temperature decreased from 34°C to 11.3°C, the total nitrogen removal rate of the DN-PN-Anammox system gradually decreased from the peak value of 1.42 kg/(m³?day) to 0.49 kg/(m³?day). At low temperatures (<20°C), when the nitrogen load (NLR) of the system is not appropriate, the fluctuation of high NH₄⁺-N concentration in the landfill leachate greatly influenced the stability of the nitrogen removal. At temperatures of 11°C–15°C, the NLR of the system is controlled below 0.5 kg/(m³?day), which can achieve stable nitrogen removal and the nitrogen removal efficiency can reach above 96%. The abundance of Candidatus Brocadia gradually increased with the decrease of temperature. Nitrosomonas, Candidatus Brocadia and Candidatus Kuenenia as the main functional microorganisms in the low temperature.     

Insights into the mechanism of the deterioration of mainstream partial nitritation/anammox under low residual ammonium

Residual ammonium is a critical parameter affecting the stability of mainstream partial nitritation/anammox (PN/A), but the underlying mechanism remains unclear. In this study, mainstream PN/A was established and operated with progressively decreasing residual ammonium. PN/A deteriorated as the residual ammonium decreased to below 5 mg/L, and this was paralleled by a significant loss in anammox activity in situ and an increasing nitrite oxidation rate. Further analysis revealed that the low-ammonium condition directly decreased anammox activity in situ via two distinct mechanisms. First, anammox bacteria were located in the inner layer of the granular sludge, and thus were disadvantageous when competing for ammonium with ammonium-oxidizing bacteria (AOB) in the outer layer. Second, the complete ammonia oxidizer (comammox) was enriched at low residual ammonium concentrations because of its high ammonium affinity. Both AOB and comammox presented kinetic advantages over anammox bacteria. At high residual ammonium concentrations, nitrite-oxidizing bacteria (NOB) were effectively suppressed, even when their maximum activity was high due to competition for nitrite with anammox bacteria. At low residual ammonium concentrations, the decrease in anammox activity in situ led to an increase in nitrite availability for nitrite oxidation, facilitating the activation of NOB despite the dissolved oxygen limitation (0.15–0.35 mg/L) for NOB persisting throughout the operation. Therefore, the deterioration of mainstream PN/A at low residual ammonium was primarily triggered by a decline in anammox activity in situ. This study provides novel insights into the optimized design of mainstream PN/As in engineering applications.     

Determination of ammonium on an integrated microchip with LED-induced fluorescence detection

A simply fabricated microfluidic device integrated with a fluorescence detection system has been developed for on-line determination of ammonium in aqueous samples. A 365-nm light-emitting diode (LED) as an excitation source and a minor band pass filter were mounted into a polydimethylsiloxane (PDMS)-based microchip for the purpose of miniaturization of the entire analytical system. The ammonium sample reacted with o-phthaldialdehyde (OPA) on-chip with sodium sulfite as reducing reagent to produce a fluorescent isoindole derivative, which can emit fluorescence signal at about 425 nm when excited at 365 nm. Effects of pH, flow rate of solutions, concentrations of OPA-reagent, phosphate and sulfite salt were investigated. The calibration curve of ammonium in the range of 0.018-1.8 μupg/mL showed a good linear relationship with R² = 0.9985, and the detection limit was (S/N = 3) 3.6 × 10^-4 μupg/mL. The relative standard deviation was 2.8% (n = 11) by calculating at 0.18 μupg/mL ammonium for repeated detection. The system was applied to determine the ammonium concentration in rain and river waters, even extent to other analytes fluorescence detection by the presented device.     

Methanogenic community dynamics in anaerobic co-digestion of fruit and vegetable waste and food waste

A lab-scale continuously-stirred tank reactor (CSTR), used for anaerobic co-digestion of fruit and vegetable waste (FVW) and food waste (FW) at different mixture ratios, was operated for 178 days at the organic loading rate of 3 kg VS (volatile solids)/(m³.day). The dynamics of the Archaeal community and the correlations between environmental variables and methanogenic community structure were analyzed by polymerase chain reactions - denaturing gradient gel electrophoresis (PCR-DGGE) and redundancy analysis (RDA), respectively. PCR-DGGE results demonstrated that the mixture ratio of FVW to FW altered the community composition of Archaea. As the FVW/FW ratio increased, Methanoculleus, Methanosaeta and Methanosarcina became the predominant methanogens in the community. Redundancy analysis results indicated that the shift of the methanogenic community was significantly correlated with the composition of acidogenic products and methane production yield. Different mixture ratios of substrates led to different compositions of intermediate metabolites, which may affect the methanogenic community. These results suggested that the analysis of microbial communities could be used to diagnose anaerobic processes.     

A flow cytometer based protocol for quantitative analysis of bloom-forming cyanobacteria (Microcystis) in lake sediments

A quantitative protocol for the rapid analysis of Microcystis cells and colonies in lake sediment was developed using a modified flow cytometer, the CytoSense. For cell enumeration, diluted sediment samples containing Microcystis were processed with sonication to disintegrate colonies into single cells. An optimized procedure suggested that 5 mg dw (dry weight)/mL dilution combined with 200 W × 2 min sonication yielded the highest counting efficiency. Under the optimized determination conditions, the quantification limit of this protocol was 3.3×10⁴ cells/g dw. For colony analysis, Microcystis were isolated from the sediment by filtration. Colony lengths measured by flow cytometry were similar to those measured by microscopy for the size range of one single cell to almost 400 μupm in length. Moreover, the relationship between colony size and cell number was determined for three Microcystis species, including Microcystis flos-aquae, M. aeruginosa and M. wessenbergii. Regression formulas were used to calculate the cell numbers in different-sized colonies. The developed protocol was applied to field sediment samples from Lake Taihu. The results indicated the potential and applicability of flow cytometry as a tool for the rapid analysis of benthic Microcystis. This study provided a new capability for the high frequency monitoring of benthic overwintering and population dynamics of this bloom-forming cyanobacterium.     

Influence and mechanism of N-(3-oxooxtanoyl)-L-homoserine lactone (C8-oxo-HSL) on biofilm behaviors at early stage

N-acyl-homoserines quenching, enzymatic quenching of bacterial quorum sensing, has recently applied to mitigate biofilm in membrane bioreactor. However, the effect of AHLs on the behavior of biofilm formation is still sparse. In this study, Pseudomonas aeruginosa biofilm was formed on ultra-filtration membrane under a series of N-(3-oxooxtanoyl)-L-homoserine lactone (C₈-oxo-HSL) concentrations. Diffusing C₈-oxo-HSL increased the growth rate of cells on biofilm where the concentration of C₈-oxo-HSL was over 10^-7 g/L. The C₈-oxo-HSL gradient had no observable influence on cell density and extracellular polymeric substances of biofilm with over 10^-7 g/L C₈-oxo-HSL. Surprisingly, 10^-11-10^-8 g/L of C₈-oxo-HSL had no effect on cell growth in liquid culture. The cell analysis demonstrated that the quorum sensing system might enhance the growth of neighboring cells in contact with surfaces into biofilm and may influence the structure and organization of biofilm.     

Seasonal trend of ambient PCDD/Fs in Tianjin City, northern China using active sampling strategy

The spatial concentrations, seasonal trends, profiles and congener pairs of ambient polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were investigated within a seasonally active sampling scheme during Jun 2008 and Jan 2009 in Tianjin City, northern China. The PCDD/F concentrations ranged 14.2-172 fg I-TEQ/m³ (average 69.3 fg I-TEQ/m³) in summer and (89.8-1.01) × 10³ fg I-TEQ/m³ (average 509 fg I-TEQ/m³) in winter, respectively, except for the E-waste dismantling site where much higher values were observed (1.04 × 10³ fg I-TEQ/m³ in summer and 7.123 × 10³ fg I-TEQ/m³ in winter). The results indicated a significantly seasonal trend with higher TEQ values in winter as compared with summer, which could be related to increased emission sources and seasonal variations of the atmospheric boundary layer height. 2,3,4,7,8-PeCDF was the dominant contributor to the total PCDD/F toxic equivalents, and 2,3,7,8-TCDD was detected at almost all the sampling sites in winter. Most of the similarly substituted PCDD/F congener pairs exhibited high correlations, suggesting that they might have similar environmental fate or sources. But different seasonal and spatial distributions of PCDD/F concentrations indicated that the emission sources might be intermittent.     

Ammonium removal pathways and microbial community in GAC-sand dual media filter in drinking water treatment

A GAC-sand dual media filter (GSF) was devised as an alternative solution for drinking water treatment plant to tackle the raw water polluted by ammonium in place of expensive ozone-GAC processes or bio-pretreatments. The ammonium removal pathways and microbial community in the GSFs were investigated. The concentrations of ammonium, nitrite and nitrate nitrogen were monitored along the filter. Total inorganic nitrogen (TIN) loss occurred during the filtration. For 1 mg ammonium removal, the TIN loss was as high as 0.35 mg, DO consumption was 3.06 mg, and alkalinity consumption was 5.55 mg. It was assumed that both nitrification and denitrification processes occur in the filters to fit the TIN loss and low DO consumption. During the filtration, nitritation, nitrification and nitritation-anaerobic ammonium oxidation processes probably occur, while traditional nitrification and denitrification and simultaneous nitrification and denitrification processes may occur. In the GSFs, Nitrosomonas and Nitrospira are likely to be involved in nitrification processes, while Novosphingobium, Comamonadaceae and Oxalobacteraceae may be involved in denitrification processes.     

Spatial distribution of archaeal and bacterial ammonia oxidizers in the littoral buffer zone of a nitrogen-rich lake

The spatial distribution and diversity of archaeal and bacterial ammonia oxidizers (AOA and AOB) were evaluated targeting amoA genes in the gradient of a littoral buffer zone which has been identified as a hot spot for N cycling. Here we found high spatial heterogeneity in the nitrification rate and abundance of ammonia oxidizers in the five sampling sites. The bacterial amoA gene was numerically dominant in most of the surface soil but decreased dramatically in deep layers. Higher nitrification potentials were detected in two sites near the land/water interface at 4.4-6.1 μg NO₂^--N/(g dry weight soil·hr), while only 1.0-1.7 μg NO₂^--N/(g dry weight soil·hr) was measured at other sites. The potential nitrification rates were proportional to the amoA gene abundance for AOB, but with no significant correlation with AOA. The NH4 + concentration was the most determinative parameter for the abundance of AOB and potential nitrification rates in this study. Higher richness in the surface layer was found in the analysis of biodiversity. Phylogenetic analysis revealed that most of the bacterial amoA sequences in surface soil were affiliated with the genus of Nitrosopira while the archaeal sequences were almost equally affiliated with Candidatus ‘Nitrososphaera gargensis’ and Candidatus ‘Nitrosocaldus yellowstonii’. The spatial distribution of AOA and AOB indicated that bacteria may play a more important role in nitrification in the littoral buffer zone of a N-rich lake.     

BT~(MT)生物膜载体对厌氧氨氧化反应器启动的影响

采用两套厌氧氨氧化反应器R1和R2,研究了BMTM生物膜载体对厌氧氨氧化工艺启动特性的影响.结果表明,R1采用UASB反应器启动厌氧氨氧化反应器,经140d运行,对氨氮和亚硝酸盐氮的去除率仅达到54.6%和58.8%,氨氮与亚硝酸盐氮去除负荷之和仅为0.09kg/(m3·d),随后,向其上部投加0.6LBMTM载体,经过26d运行,氨氮及亚硝酸盐氮去除率分别迅速提升至92.5%和97.4%,R1的启动速度较之前有明显提高;R2采用BMTM载体启动上流式填料床生物膜反应器厌氧氨氧化工艺,经过83d的运行,氨氮及亚硝酸盐氮去除率分别达到83.6%和89.4%,氨氮与亚硝酸盐氮去除负荷之和达0.22kg/(m3·d),启动速度较R1大幅提高.     

部分亚硝化-厌氧氨氧化耦合工艺处理污泥脱水液

在缺氧滤床+好氧悬浮填料生物膜工艺中实现部分亚硝化,然后进行厌氧氨氧化(ANAMMOX),考察其对高含氮、低C/N污泥脱水液的处理能力.结果表明,亚硝化反应器在15~29℃、DO 6~9mg/L条件下,通过综合调控进水氨氮负荷(ALR)、进水碱度/氨氮、水力停留时间(HRT)等运行参数,可以调节出水(NO2--N)/(NH4+-N)的比率,能够较好地实现部分亚硝化反应以完成厌氧氨氧化.当进水ALR为1.16kg/(m3·d),进水碱度/氨氮为5.1时,出水(NO2--N)/(NH4+-N)在1.2左右,(NO2--N)/(NOx--N)大于90%,进入ANAMMOX反应器的氮物质去除率达到83.8%.     

AOB与AnAOB在不同生物填料上挂膜效果的研究

针对厌氧氨氧化工艺运行过程中污泥流失严重与启动时间长等问题,本研究通过对多种市售生物填料的挂膜实验,筛选适合好氧氨氧化菌与厌氧氨氧化菌挂膜的生物填料.结果表明,当活性污泥中的好氧氨氧化菌菌属为Nitrosomonas时,AQ1聚氨酯立方体填料最适合其挂膜,挂膜成熟时好氧氨氧化速率可以达到（0.81±0.08）mg N/（L·h）,挂膜生物量为（0.87±0.14）mg VSS.而更适合厌氧氨氧化菌（Candidatus Kuenenia）挂膜的生物填料为K3环形填料,材质为聚乙烯或者聚丙烯.当厌氧氨氧化菌挂膜成熟时,其厌氧氨氧化速率可以达到（3.27±0.10）mg N/（L·h）,挂膜生物量为（2.74±0.40）mg VSS.厌氧氨氧化菌在K3型填料上的挂膜要优于好氧氨氧化菌,其原因是厌氧氨氧化菌分泌的胞外聚合物含量要高于好氧氨氧化菌,而胞外聚合物是形成生物膜的重要因素.     

Anammox transited from denitrification in upflow biofilm reactor

Anammox was successfully transited from heterotrophic denitrification and autotrophic denitrification in two upflow biofilm reactors, respectively. The results showed that the volumetric loading rate and nitrogen removal efficiency in the reactor transited from heterotrophic denitrification were higher than that in its counterpart. When the hydraulic retention time was 12 h or so, the total nitrogen loading rate was about 0.609 kg N/(m^3 .d), and the effluent ammonia and nitrite concentrations were less than 8.5 mg/L and 2.5 mg/L,respectively. The upflow anammox biofilm reactor was capable of keeping and accumulating the slow-growing bacteria efficiently. During operation of the reactor, the biomass color was gradually turned from brownish to red, and the ratio of ammonia consumption, nitrite consumption and nitrate production approached the theoretical one. These changes could be used as an indicator for working state of the reactor.     

Performance and microbial response during the fast reactivation of Anammox system by hydrodynamic stress control

Anaerobic ammonium oxidation （Anammox） has become a promising method for biological nitrogen removal. However, this biotechnology application is always limited due to the low growth rate and biomass yield of Anammox bacteria. This study investigated the process of fast reactivation of an Anammox consortium idled for 2 years uia hydrodynamic stress control. The results showed that the Anammox system was efficiently and quickly reactivated by shortening of the hydraulic retention time （I-IRT） of the reactor from 12 to 6 hr within 68 days of operation. Moreover, at a 4-hr HRT with an influent total nitrogen loading rate of 1.2 kg N/（m3.day）, the reactor maintained high biological performance with an ammonium removal loading rate of 0.52 kg N/（m3.day） and a nitrite removal rate of 0.59 kg N/（m3.day）. In the reactivated Anammox reaction, the stoichiometric coefficients of NH4-N to NOE-N and NH4-N to NO4-N were 1：1.04± 0.08 and 1：0.31 ± 0.03, respectively. The specific Anammox activity and hydrazine oxidoreductase activity, both of which represent the degree of Anammox bacteria present, increased as the hydrodynamic stress increased and were maximally （125.38 ± 3.01 mg N/（g VSS.day） and 339.42 ± 6.83 μmol/（min.g VSS）, respectively） at 4-hr HRT. Microbial response analysis showed that the dominant microbial community was obviously shifted and the dominance of Anammox bacteria was enhanced durinR the hydrodynamic selection.     

Performance of Anammox granular sludge bed reactor started up with nitrifying granular sludge

IntroductionTheanaerobicammoniaoxidation(Anammox)isanovelbiologicalreactionthatproducesmolecularnitrogenwithammoniaaselectrondonorandnitriteaselectronacceptor,respectively (vandeGraff,1995 ;1996 ) .Anammoxprocesshasbeenshowntobeapromisingwayofremovingnitr…     

An innovative membrane bioreactor and packed-bed biofilm reactor combined system for shortcut nitrification-denitrification

An innovative shortcut biological nitrogen removal system, consisting of an aerobic submerged membrane bioreactor (MBR) and an anaerobic packed-bed biofilm reactor (PBBR), was evaluated for treating high strength ammonium-bearing wastewater. The system was seeded with enriched ammonia-oxidizing bacteria (AOB) and operated without sludge purge with a decreased hydraulic retention time (HRT) through three phases. The MBR was successful in both maintaining nitrite ratio over 0.95 and nitrification efficiency higher than 98% at a HRT of 24 h. The PBBR showed satisfactory denitrification efficiency with very low effluent nitrite and nitrate concentration (both below 3 mg/L). By examining the nitrification activity of microorganism, it was found that the specific ammonium oxidization rate (SAOR) increased from 0.17 to 0.51 g N/(g VSS·d) and then decreased to 0.22 g N/(g VSS·d) at the last phase, which resulted from the accumulation of extracellular polymers substances (EPS) and inert matters enwrapped around the zoogloea. In contrast, the average specific nitrite oxidization rate (SNOR) is 0.002 g N/(g VSS·d), only 1% of SAOR. Because very little Nitrobactor has been detected by fluorescence in situ hybridization (FISH), it is confirmed that the stability of high nitrite accumulation in MBR is caused by a large amount of AOB.     

序批式生物膜CANON工艺的运行与温度的影响

通过序批式生物膜反应器,研究了CANON工艺的运行规律,并研究了温度对CANON工艺的影响.研究发现,通过序批式生物膜反应器反应器可以成功实现CANON工艺,TN去除率可达到88.3%,TN去除负荷达到0.52kg/(m3·d).系统中的NO2--N变化规律可以分为浓度上升期,浓度稳定期与浓度下降期.pH值先降低后升高的拐点与DO突跃的拐点均可作为反应结束的指示参数,DO拐点比pH值拐点出现要提前.在26~35℃之间,可以获得较好的硝化效果与TN去除效果,温度降低到20℃时,厌氧氨氧化性能开始受到显著影响,造成反应器中大量NO2--N积累,而硝化效果在15℃才受到显著影响.反应器中厌氧氨氧化细菌的最适温度约为30℃.     

生物膜全自养脱氮过程及强化机理研究进展

在限制溶氧的条件下,生物膜表层的好氧氨氧化菌将氨氧化为亚硝酸盐,并传递到生物膜内层缺氧区,厌氧氨氧化菌将氨和亚硝酸盐同步去除。根据生物膜内好氧氨氧化菌和厌氧氨氧化菌协作共生开发出的全自养脱氮工艺在生物转盘、SBR及填料床等反应器内实现,大大节省了碳源和曝气量。微量NO2对好氧氨氧化和厌氧氨氧化过程有明显强化作用,能显著提高基质降解速率、微生物生长速率和细胞密度。数学模型对废水处理工艺开发和运行具有重要意义,通过数学模型对微生物活性、分布以及脱氮过程的主要影响因素进行模拟研究,为全自养脱氮工艺的运行管理和优化控制打下基础。     

部分反硝化耦合厌氧氨氧化IFAS系统脱氮性能

采用部分反硝化活性污泥耦合厌氧氨氧化生物膜处理低碳氮比废水(C/TN=1.63),考察生物膜-活性污泥复合系统(IFAS)进行部分反硝化耦合厌氧氨氧化(PD/A)处理低碳氮比废水的可行性及其耦合后两相中功能菌活性与菌群分布变化规律.结果显示,系统耦合运行期间,出水TN为(5.07±0.2)mg/L,去除率为(90.7±0.1)%,厌氧氨氧化途径对TN去除的贡献率高达(86.61±3.4)%;固着相对厌氧氨氧化活性的贡献率为100%,悬浮相上,μ(NO₃^--N)占比为99.32%,μ(NO₂^--N)占比为99.22%;与耦合前相比,悬浮相中硝酸盐还原酶(Nar)活性由(0.43±0.05)μmol/(mg protein·min)增加至(0.49±0.09)μmol/(mg protein·min),亚硝酸盐转化率明显升高[(70±2.2)%~(90.01±2.3)%];Illumina MiSeq结果显示,固着相上的优势菌属为Candidatus_Brocadia,且耦合前后丰度无明显变化(33.61%~33.43%),悬浮相上反硝化菌属Prosthecobacter,Ferruginibacter,OLB8丰度增加.以上结果表明,在IFAS系统中可以实现稳定的PD/A协同脱氮,耦合后部分反硝化由悬浮相主导,厌氧氨氧化由固着相主导,厌氧氨氧化菌(AnAOB)与反硝化菌对NO₂^­--N的竞争强化了悬浮相部分反硝化能力.