厌氧氨氧化菌接种污泥的选择培养过程研究

厌氧氨氧化菌的2种不同接种污泥培养实验表明,厌氧消化污泥和好氧硝化污泥均可成功启动厌氧氨氧化过程.接种厌氧消化污泥比好氧硝化污泥培养的厌氧氨氧化菌启动快,但后者去除效果较好.接种好氧硝化污泥的反应器的厌氧氨氧化速率随着氨氮基质进水浓度的增加呈线性增加.进水氨氮浓度为280 mg/L时的氨氮平均去除率达91%;而接种厌氧消化污泥的相应氨氮平均去除率仅为52%.厌氧氨氧化过程以接种好氧硝化污泥来启动为宜.     

不同污泥源条件下ANAMMOX启动及其与反硝化协同脱氮对比

采用城市生活污水配水同时启动两组ASBR,R1接种好氧硝化污泥,R2按2∶1混合接种短程硝化和厌氧氨氧化污泥,研究2个ANAMMOX反应器启动的可行性及其差异。实验结果表明,R1和R2均可成功启动ANAMMOX,R1需130 d,R2仅需73 d;稳定期R1和R2反应器NH4+-N、NO2--N和TN去除率分别达95.30%、91.30%、76.28%和96.2%、98.3%、90.1%,且周期内NH4+-N、NO2--N和NO3--N降解规律相似;R1和R2反应器发生的主要反应为厌氧氨氧化,但同时存在反硝化作用;2组反应器稳定运行后污泥颜色、形态及微生物组成相似,经SEM观察多为球状菌。     

ASBR厌氧氨氧化反应器的快速启动及脱氮原理分析

以城市生活污水为基本水质进行配水,采用ASBR研究了厌氧氨氧化反应器的快速启动过程及脱氮性能。实验条件如下:T为(35±1)℃、HRT为24 h、pH为7.2～7.5,进水NH4+-N、NO2--N浓度为40～160 mg/L,TN负荷为0.08～0.34 kg TN/(m3.d),按2∶1比例混合接种好氧短程硝化污泥和厌氧氨氧化污泥,经49 d运行成功启动厌氧氨氧化反应器,并实现稳定运行。实验结果表明:稳定运行期NH4+-N、NO2--N去除率分别达96%和98%;NH4+-N、NO2--N去除量与NO3--N生成量比值为1∶1.05∶0.29,较为接近理论值;成功启动的反应器出水pH高于进水;系统TN去除率平均值为79.7%;反应器内存在反硝化与厌氧氨氧化的协同作用,实现了部分COD去除;污泥由深棕色絮状变成红褐色颗粒状,经SEM扫描电镜观察污泥菌群种类单一,多为球状菌,有漏斗状缺口,具有典型氨氧化菌形态特征。     

不同泥源对厌氧氨氧化反应器启动的影响

采用2套上流式生物膜反应器,分别接种少量厌氧氨氧化污泥和大量硝化污泥,考察其对厌氧氨氧化反应器启动的影响。污泥接种入反应器后,测得接种厌氧氨氧化污泥的反应器(R1)内MLSS为0.22 g/L,另一个反应器(R2)MLSS为2.7 g/L。与直接接种厌氧氨氧化污泥相比,R1经过72 d的运行才显现出厌氧氨氧化特性。经过114 d的培养,前者氮去除速率由0.23 kg/(m3.d)提升到5.29 kg/(m3.d),总氮去除率大于89%;R2的氮去除速率由0.01 kg/(m3.d)提升到1.1 kg/(m3.d),总氮去除率大于84.6%。说明普通污泥启动需要一个较长的筛选过程,直接接种少量的厌氧氨氧化污泥比接种普通的污泥能够更快启动厌氧氨氧化反应器。     

城市污水条件下ASBR厌氧氨氧化的启动与脱氮性能

采用ASBR在城市污水条件下进行厌氧氨氧化的启动与脱氮性能研究。实验接种好氧硝化污泥,在温度为(35±1)℃、HRT为24 h、pH为7.3～8.5的条件下经过130 d的培养,成功启动了厌氧氨氧化反应。实验结果表明,厌氧氨氧化反应稳定运行时,TN容积负荷平均为0.179 kg/(m3.d),NH4+-N、NO2--N和TN去除率分别达到了95.30%、91.30%和76.28%。启动期和稳定运行期NH4+-N、NO2--N去除量和NO3--N生成量的比值分别达到1∶1.54∶0.25和1∶1.27∶0.27;稳定运行期进出水pH差值由启动时的0.85下降到0.24。启动期反硝化和厌氧氨氧化反应同时存在而在稳定运行期厌氧氨氧化发展成为主导反应。MLSS和MLVSS/MLSS先减少后增加,反映了启动过程中硝化细菌、反硝化细菌的衰亡和厌氧氨氧化菌逐渐富集的过程,这与反应器的宏观运行效果相一致。     

不同污泥源厌氧氨氧化污泥的比较

采用厌氧序批式反应器,以好氧硝化污泥和厌氧颗粒污泥为污泥源,通过对氨氮、亚硝酸盐氮、pH等指标的监测和数据分析、污泥颜色变化和菌落电镜照片的观察,研究2种不同污泥源厌氧氨氧化污泥的差异.结果表明,不同污泥源厌氧氨氧化污泥的颜色不同,污泥中具有厌氧氨氧化作用的优势菌不同;由厌氧颗粒污泥为污泥源培养出的厌氧氨氧化污泥具有较高的厌氧氨氧化活性.     

改良型UASB厌氧氨氧化反应器运行效能对比

为探究组合启动模式实现厌氧氨氧化反应器高效启动和稳定运行的可行性,分别采用接种短程硝化污泥结合提高进水基质(A)和接种厌氧氨氧化污泥结合缩短水力停留时间(B)2种组合方式启动改良型UASB厌氧氨氧化反应器,对反应器启动效果进行研究,并通过改变进水基质比和低温冲击探究启动成功后的反应器性能。结果表明:A反应器启动成功时的总氮去除负荷(NRR)为0.520 kg·(m~3·d)~(-1)、亚硝化单胞菌Nitrosomonas相对丰度大幅下降、主要厌氧氨氧化菌属由Candidatus Kuenenia转化为Candidatus Brocadia;而B反应器NRR达到1.950 kg·(m~3·d)~(-1)、Candidatus Kuenenia始终为优势菌属。随着进水基质比的提高,B反应器的NRR和上升幅度始终高于A反应器,具有更强的抗负荷能力。当温度由35℃下降至15℃时,A和B反应器污泥对基质的降解速率分别下降92.94%和81.38%;温度恢复至35℃后,A反应器污泥降解速率的回升率大于B反应器污泥。因此,接种厌氧氨氧化污泥和缩短水力停留时间的组合方式更有利于改良型UASB厌氧氨氧化反应器的高效启动和稳定运行。     

基于SBR反应器的ANAMMOX工艺的启动运行

采用SBR反应器,接种好氧硝化污泥,在142 d内于较高负荷下成功启动了厌氧氨氧化反应器.反应器总氮容积负荷(以N计)为0.43 kg/m3·d,总氮去除率最高达到93.3%,平均为80.5%;氨氮和亚硝酸盐氮的去除率最高达到93.9%和99.8%,平均去除率为81.2%和85.7%.在稳定运行阶段,氨氮去除量、亚硝酸盐氮去除量、硝酸盐氮生成量三者之间的比值为1:1.38:0.18.反应器启动过程中,出水、进水pH差值的变化趋势由负到正,然后稳定在一定范围内;且污泥性状有较大变化,污泥中微生物所占比率有所提高,整个反应器中适应厌氧氨氧化运行方式的菌种增殖较快.     

基于SBBR的单级自养脱氮快速启动

以普通活性污泥为接种污泥,采用人工配制无机氨氮废水进行单级自养脱氮工艺快速启动研究。启动过程经历了污泥适应期、部分短程硝化选择期以及单级自养脱氮实现期3个阶段。经过29 d的培养驯化,通过控制游离氨的方法实现了部分短程硝化。当出水中亚硝酸盐积累率达到60%左右时,立即将序批式生物膜反应器(SBBR)由连续曝气改为间歇曝气,间歇曝气使得厌氧氨氧化菌(AAOB)的富集与亚硝酸氧化菌(NOB)的淘汰同时进行,并且避免了高浓度亚硝酸盐对AAOB的抑制作用,从而实现了单级自养脱氮的快速启动。实验仅用50 d成功启动了SBBR单级自养脱氮工艺,总氮容积去除负荷达到0.173 kg N/(m3·d),氨氮的平均去除率达到98.68%,总氮的平均去除率达到80.87%。成功启动之后,反应器内只有少量的悬浮污泥,大部分的污泥都附着在填料上,污泥颜色呈褐色,而反应器内壁及出水管上附着的污泥呈浅砖红色,表明反应器内富集了大量的AAOB。     

pH和C:N对厌氧氨氧化耦合短程反硝化脱氮性能的影响

以低DO和逐渐降低亚硝态氮浓度的方式运行厌氧氨氧化(ANAMMOX)上流式污泥床(AUASB)反应器,第57天实现稳定运行,氨氮去除率保持在85%以上,采用高通量测序技术分析发现ANAMMOX污泥中主要功能菌转变为Candidatus kuenenia,丰度为8.85%,好氧氨氧化菌(AOB)Nitrosomonas的丰度为1.48%,短程反硝化菌Thauera丰度为0.66%。将AUASB反应器中污泥接种在ANAMMOX序批式反应器(ASBR)后,通过外加有机碳源的方式使ASBR拥有了短程反硝化的能力,然后研究了不同C:N(质量比)和不同pH条件下亚硝态氮积累情况和氮素转化特性,结果表明:当C:N为3.2:1.0时,亚硝态氮积累率最高为84%,但有部分亚硝态氮被还原。当pH为7.8时,ANAMMOX菌活性最高,氨氮去除率为95%。扫描电子显微镜(SEM)观察到ANAMMOX菌为球状,短程反硝化菌为短杆状。     

有机碳源条件下厌氧氨氧化SBBR中的微生物多样性

研究有机碳源对SBBR厌氧氨氧化菌群等微生物的影响。采用16S rDNA序列与PCR-DGGE分析技术相结合的方法,对稳定运行的反应器内的活性污泥和生物膜样品,进行细菌多样性图谱分析,同时采用巢式PCR-DGGE技术对浮霉状菌属(Planctomycetes)细菌进行分析。结果表明,在有机碳源反应系统细菌条带数和多样性指数均高于无机系统,与活性污泥相比,生物膜表尤为明显。当进水不含有机碳源时,氨氧化细菌(ammonia oxidizing bacteria,AOB),厌氧氨氧化菌(anaerobic ammonia oxidizing bacteria,ANAMMOX)为优势功能菌;当进水含有机碳源时,系统中存在的AOB以亚硝化单胞菌(Nitrosomonas sp.)为优势菌群,同时存在反硝化菌,如索氏菌(Thauera sp.)以及厌氧氨氧化菌,它们共同作用完成N的去除。此外,与无机碳源系统相比,有机碳源的存在,有利于浮霉状菌的积累,但压缩了ANAMMOX的生存空间。本研究可为厌氧氨氧化工艺处理低C/N比有机废水提供了理论依据。     

Anammox enrichment from different conventional sludges

Three sets of sequencing batch reactor (SBR) were used for Anammox enrichment from conventional sludges including upflow anaerobic sludge blanket, activated sludge, and anaerobic digestion sludge. After four months of operation, the Anammox activity occurred in all reactors allowing continuous removal of ammonium and nitrite. The morphology of the cultivated Anammox sludge was observed using scanning electron microscope. The photographs showed that the obtained culture was mostly spherical in shape, presumably Anammox culture. There were also filamentous-like bacteria co-existing in the system. Fluorescence in situ hybridization (FISH) analysis using 16S rRNA targeting oligonucleotide probes PLA46 and Amx820 showed that the dominant population developed in all SBRs was hybridized with both PLA46 and Amx820 gene probes. It means that the cultivated biomass in all SBRs was classified in the group of Planctomycetales bacteria with respect to the anaerobic ammonium-oxidizing bacteria, Candidatus Brocadia anammoxidans and Candidatus Kuenenia stuttgartiensis. Numerous time sequences were tested in this experiment. The shortest workable reaction time was found in the range from 5 to 7 h. Good quiescence of sludge was obtained at 30 min of settle period followed by a discharge period of 15 min. A long-term performance showed a near perfect removal of nitrite based on the influent NO2(-)-N concentration of 50-70 mg l(-1). The maximum ammonia removal efficiency was 80% with the influent NH4(+)-N concentration of 40-60 mg l(-1). It is, therefore, concluded that Anammox cultivation from conventional sludges was highly possible under control environment within four months.     

硝化颗粒污泥的快速培养及其硝化特性分析

以好氧颗粒污泥接种小试柱形SBR，采用自配无机氨氮废水为进水，在中温（28～30℃）条件下通过逐步提升进水NH4^＋-N浓度（100～650mg／L）和缩短水力停留时间（8～4h）快速培养硝化颗粒污泥。实验结果证实，以好氧颗粒污泥接种可以促使硝化颗粒污泥快速形成，36d时粒径〉0．21mm的颗粒污泥占总数的93％，颗粒污泥NH4-N比去除速率为50．53mgNH4^＋-N／（gSS·h）。硝化颗粒污泥具有良好的短程硝化性能，亚硝酸盐产生速率和累积率分别保持在3．3kgNO2-N／（m^3·d）和85％以上。反应初期高FA和反应末期高FNA的共同抑制是该研究中实现和维持稳定短程硝化的关键因素。     

Biotransformation of estrogens in nitrifying activated sludge under aerobic and alternating anoxic/aerobic conditions.

Natural and synthetic estrogens present in municipal wastewater can be biodegraded during treatment, particularly in activated sludge. The objective was to assess the extent of transformation of 17-beta-estradiol (E2) and 17-alpha-ethinylestradiol (EE2) by nitrifying activated sludge and evaluate potential relationships between availability of oxygen, nitrification rate, and estrogen removal. For each batch experiment, two reactors were set up--aerobic and alternating anoxic/aerobic-which were then amended with E2 and EE2 from methanolic stock solutions. The EE2 was persistent under anoxic conditions; under aerobic conditions, the observed level of its removal was 22%. The E2 was readily converted to estrone (El)--faster under aerobic (nitrifying) than anoxic (denitrifying) conditions. During the initial anoxic conditions, a metabolite consistent with 17-alpha-estradiol transiently accumulated and was subsequently removed when the reactor was aerated. Higher removal rates of estrogens were associated with higher nitrification rates, which supports the contention that the nitrifying biomass was responsible for their removal.     

Improving nitrogen removal in two modified decentralized wastewater systems

Efficient nutrient removal in decentralized wastewater treatment systems is a challenging task. To improve the removal of organic matter and nitrogen from wastewater, two types of bioreactors using membrane-aerated biofilm reactor (MABR) and microbial fuel cell (MFC) techniques were evaluated. During more than 250 days of continuous-flow reactor operation, both reactors showed consistently high chemical oxygen demand removal (>86%). At an influent ammonium-nitrogen (NH4(+)-N) concentration of 30 mg N/L, the average effluent NH4(+)-N concentrations were 6.2 and 0.5 mg N/L for the MABR and MFC reactor, respectively, while the effluent nitrate-nitrogen (NO3(-)-N) concentrations were 5.4 mg/ L in the MABR and 19.2 mg/L in the MFC-based reactor. The overall total inorganic nitrogen removal efficiencies were 64% and 36% for the MABR and MFC reactor, respectively. At the measured dissolved oxygen concentrations of 5.2 and 0.23 mg/L in the aerobic/anoxic zone of the MFC and MABR, respectively, a specific oxygen uptake rate of 0.1 g O2/g VSS-d, resulting from ammonia oxidation, was detected in the settled sludge of the MFC, while no nitrifying activity of the sludge from the MABR was detected. Molecular microbial analysis demonstrated a link between the bacterial community structure and nitrifying activity. The relatively high abundance of Nitrosomonas europaea was associated with its detectable nitrification activity in the settled sludge of the MFC. The results suggest that MABR and MFC techniques have the potential to improve organic and nitrogen removal in decentralized wastewater systems.     

Stoichiometric and molecular evidence for the enrichment of anaerobic ammonium oxidizing bacteria from wastewater treatment plant sludge samples

Anammox enrichments were readily developed from seven municipal wastewater treatment plants (WWTPs) sludge, but not with methanogenic granular sludge from two agro-industrial WWTPs. Only 50 d was required for the first evidence of anammox activity from a return activated sludge obtained from a WWTP operated for nutrient removal. The molar ratios of nitrite and ammonium consumption of approximately 1.32 as well as nitrate and dinitrogen gas product ratios of approximately 0.095 provided evidence of the anammox reaction. The presence of anammox was confirmed by polymerase chain reaction (PCR) using primer sets (PLA46F and AMX820R) specific for anammox bacteria. The 16S rRNA gene fragment of anammox bacteria was detected in seven enrichment cultures (ECs) with demonstrated anammox activity but not in the original inocula from which the ECs were derived and also not in the two methanogenic sludge samples, which indicates the PCR predicted the anammox activity. Two genera, Brocadia and Kuenenia, were successfully identified as the Planctomycetes occurring in the clone libraries of successful anammox enrichments. Brocadia dominated in cultures that were respiked extensively; whereas Kuenenia predominated in cultures that were less aggressively respiked. These findings indicate that respiking management may play an important role on selecting the genus of anammox bacteria. The batch enrichment results clearly illustrate that anammox can be readily enriched from municipal sludge from a wide variety of process operations at WWTPs.     

Production of polyhydroxyalkanoate during treatment of low-phosphorus-content wastewater.

To evaluate whether poly-beta-hydroxyalkanoate (PHA) production and wastewater treatment could be combined in a single biological process, a bench-scale sequencing batch reactor was operated with sequential anaerobic and aerobic stages and removal of excess sludge at different stages of treatment. The reactor treated synthetic wastewater with a high organic and low nutrient content, simulating industrial wastewater. Chemical oxygen demand removal efficiency was more than 90% in all cases. Poly-beta-hydroxyalkanoate accumulation was significant, although it did not appear to be induced by oxygen limitations during the anaerobic stage. Sphaerotilus natans was apparently the dominant PHA-accumulating organism at the end of each reactor run and corresponded to a PHA accumulation of 16 to 20% of the total dry cell mass. Before S. natans dominated the reactors, PHA accumulation was approximately 17% when biomass was removed at the end of the aerobic stage and 6.6% when sludge removal also occurred during the anaerobic stage.     

培养方式对废水脱氮与沼气脱硫污泥驯化影响

实验研究了底物、接种污泥和微生物生长方式对猪场废水脱氮和沼气脱硫耦联污泥驯化及活性恢复的影响,以解决快速富集培养废水脱氮与沼气脱硫微生物的问题。研究发现,就脱氮脱硫均达到60%的时间而言,接种厌氧污泥反应器为9 d,比接种好氧污泥反应器(18 d)和不接种污泥加填料反应器(21 d)更短。以含氮含硫废水为底物驯化时,接种厌氧污泥更有利于脱氮脱硫污泥的驯化;而同为接种好氧污泥时,以含氮含硫废水为底物的驯化方式更有利于脱氮脱硫污泥的驯化。污泥活性恢复实验中,以含氮废水+沼气(H2S)为底物培养驯化的污泥,硫转化活性恢复所用的时间为15 d,比含氮含硫废水为底物驯化污泥的活性恢复时间更长。