一株好氧反硝化茵的分离鉴定及其混合应用特性研究

采用溴百里酚（BTB）鉴定培养基和稀释平板法从南京市某市政污水处理厂曝气池污水样本中分离筛选得到1株好氧反硝化细菌，经16SrDNA序列同源性比较和系统发育分析初步鉴定为反硝化产碱杆菌（Alcaligenes denitrificns），并将其命名为菌株BMB—N6。研究了菌株BMB—N6在不同浓度亚硝态氮条件下的反硝化能力，运用正交试验设计探讨了该菌株最适的好氧反硝化条件，并且在实验室和大田条件下分别考察了菌株BMB—N6与蛋白质降解菌BMB-LA和氨氮脱除菌BMB—HKF复配形成的混合菌制剂的反硝化能力。结果表明，菌株BMB—N6在8h内对亚硝态氮的去除率可达94％，其最适亚硝态氮去除条件为摇床转速50r·min^-1，C／N比值4，pH6，温度35℃。在实验室条件下以菌株BMB-N6为基础制成的混合菌制剂在12h内可去除90％的亚硝态氮，在大田应用中7d内可去除80％的亚硝态氮。     

1株贫营养好氧反硝化菌的分离鉴定及其脱氮特性

从水库底泥样品中,以硝酸盐为唯一氮源进行驯化、分离筛选出1株能在贫营养及好氧条件下进行高效反硝化的菌株PY8,经过电镜形态学观察、生理生化和16S rDNA序列分析,并基于16SrDNA序列结果,构建了该菌株的系统发育树,最终确定菌株PY8为根瘤菌Rhizobiumsp.。考察了初始pH值、温度、C/N、初始硝酸钠质量浓度、投菌量对菌株PY8硝酸盐还原活性的影响,以及该菌株的异养硝化性能。结果表明,在pH6.0～10.0,温度25～30℃,C/N1.0～9.0,初始硝酸钠质量浓度0.01～0.50g·L-1,投菌量1%～15%时,菌株PY8培养72h后的硝氮去除率可达到95%以上。另外,该菌株具有同时硝化-反硝化作用,在培养过程中氨氮去除率可达到58%左右。实验结果表明,菌株PY8在微污染水体生物脱氮领域中具有很大的应用潜力。     

一株贫营养异养硝化-好氧反硝化菌的筛选及脱氮特性

为了探究并优化菌剂应用于微污染水源水体修复的机制和条件,主要针对水库沉积物内筛选出的贫营养好氧反硝化菌进行了菌种鉴定及脱氮特性研究,考察菌株在不同环境条件下的脱氮效果,明确了该菌株的最适宜生长条件,并基于水库水体中贫营养条件对菌株进行水源水库原水的驯化培养试验研究,以期实现该菌株对微污染水源水库原水中氮源污染物的脱除,为原位投菌技术实际工程应用提供理论依据。从微污染水源水库沉积物中驯化筛分出一株高效异养硝化-好氧反硝化菌A14,通过扫描电镜观察、生理生化特征、16S rRNA基因测序和Biolog GenⅢ鉴定,确定该菌株为革兰氏阴性短杆菌,鉴定为皮特不动杆菌（Acinetobacter pittii）。在好氧条件下,菌株细胞内表达反硝化功能基因napA,以NO3-为唯一氮源进行反硝化作用时,36 h时NO3-去除率为78.89%。以NH4+为唯一氮源时,48 h NH4+去除率为95.25%,TN去除率达80.42%,TOC去除率达98.30%,表明该菌株具有异养硝化-好氧反硝化特性。在改变环境条件过程中,该菌株在以乙酸钠为碳源,温度为30℃,C/N为12,pH为7,接种量为10%时,NO3-去除率最高为86.62%,并且在10℃下脱氮率达到40.18%。在水源水库原水脱氮实验中,接种处理TN去除率为50.95%,NO3-去除率为80.25%。结果表明,菌株A14在微污染水源水体菌剂脱氮修复中具有良好的应用潜力。     

一株好氧反硝化菌的分离鉴定及其混合应用特性研究

采用溴百里酚(BTB)鉴定培养基和稀释平板法从南京市某市政污水处理厂曝气池污水样本中分离筛选得到1株好氧反硝化细菌,经16S rDNA序列同源性比较和系统发育分析初步鉴定为反硝化产碱杆菌(Alcaligenes denitrificans),并将其命名为菌株BMB-N6.研究了菌株BMB-N6在不同浓度亚硝态氮条件下的反硝化能力,运用正交试验设计探讨了该菌株最适的好氧反硝化条件,并且在实验室和大田条件下分别考察了菌株BMB-N6与蛋白质降解菌BMB-LA和氨氮脱除菌BMB-HKF复配形成的混合菌制剂的反硝化能力.结果表明,菌株BMB-N6在8 h内对亚硝态氮的去除率可达94%,其最适亚硝态氮去除条件为摇床转速50 r·min-1,C/N比值4,pH 6,温度35 ℃.在实验室条件下以菌株BMB-N6为基础制成的混合菌制剂在12 h内可去除90%的亚硝态氮,在大田应用中7 d内可去除80%的亚硝态氮.     

一株异养硝化-好氧反硝化细菌的分离鉴定及其脱氮性能研究

与传统脱氮菌相比,异养硝化-好氧反硝化菌在脱氮方面具有较大优势并受到广泛关注。以乙酰胺为唯一氮源从活性污泥中分离得到1株脱氮性能较高的异养硝化-好氧反硝化细菌,命名为Y1。经形态观察、生理生化特征和16S rRNA分析后鉴定为Acinetobaterjohnsonii(约氏不动杆菌),革兰氏染色结果为阴性。对Y1菌株进行生理生化鉴定试验,结果显示Y1对吲哚、柠檬酸盐、硫化氢和接触酶的反应呈阳性,表明该菌株能良好的利用以上物质;而甲基红、葡萄糖发酵、蔗糖发酵、明胶液化、淀粉水解、氧化酶、尿素酶试验结果呈阴性,表明该菌株不能很好的利用以上物质。为了检测Y1菌株的脱氮性能,将其分别置于异养硝化培养基和好氧反硝化培养基进行培养,在108 h内,接种Y1菌株的异养硝化培养基中的氨氮去除率约为66.9%,去除速率达0.53 mg.L~(-1)·h~(-1),硝氮去除率约为100%,去除速率达0.10 mg·L~(-1)·h~(-1);在84 h内,接种Y1菌株的好氧反硝化培养基中的硝氮去除率约为69.7%,去除速率达0.74 mg·L~(-1)·h~(-1),上述结果表明Y1菌株的脱氮性能较高。为了进一步研究该菌株的生长需求,保持其它条件不变的情况下,将其分别置于不同碳源和氮源下进行培养,结果表明,菌株Y1在琥珀酸钠为唯一碳源时的生长速率、异养硝化和好氧反硝化性能最好,并且利用无机氮源的能力比有机氮源能力强。     

养殖水体复合功能菌的分离及其性能

针对养殖水体中因氨态氮、硫化氢和小分子有机酸富营养化引起的污染问题,分离筛选出硝化细菌、反硝化细菌、光合细菌、硫化细菌和生物絮凝菌等具有不同生理功能的污染物治理菌株,经优化配伍制备出性能优良的复合功能菌,结果表明:硝化细菌对氨态氮的去除率达97.8%,亚硝态氮的去除率达95.7%,反硝化细菌对硝态氮的去除率为96.4%,光合细菌和硫化细菌对硫化氢的去除率为55%,微生物絮凝菌的絮凝效率为83%;复合功能菌对CODCr、NH4+-N,总氮、硫化物的去除率分别可达94.3%,89.6%,88.7%和71.3%。     

一株轻度嗜盐反硝化菌的分离鉴定及特性

从处理高盐度废水的成熟活性污泥中分离筛选得到1株轻度嗜盐反硝化细菌YL-1.通过对该菌株的形态观察、生理生化实验以及16SrDNA基因序列分析,确定该菌株为迪茨氏菌(Dietziasp.).盐度影响实验表明,该菌株能在盐度为0～10%的培养液中生长,盐度为3%时D600nm达到最大,为2.002;当初始硝态氮浓度为120mg/L左右时,DO值对该菌株的反硝化效果几乎没有影响,NOX——N去除率均在90%以上,表明该菌为好氧反硝化菌;适于在碱性环境中生长,最适pH为7.5～8.5;能利用乙酸钠、蔗糖、葡萄糖、柠檬酸钠、丁二酸钠为碳源进行反硝化,以蔗糖为碳源时反硝化效果最好,NOX——N去除率达到99.8%;在25～30℃的温度范围内反硝化效果较好,30℃时菌株的NOX——N去除率超过90%.该菌在一定盐度下具有较好的反硝化作用,能为高盐度废水的生物处理提供一定的参考价值.     

水库贫营养异养硝化-好氧反硝化菌Sxf14的脱氮特性

为利用生物强化法降低微污染源水中的氮素,从水库沉积物中筛选到一株好氧反硝化细菌Sx f14.通过扫描电镜和16S r RNA序列分析,鉴定其为不动杆菌属(Acinetobacter sp.),命名为Acinetobacter sp.Sxf14.同时,对该菌株脱氮特性进行研究,并将其接种到C/N(总有机碳与总氮的比值)为1.2的微污染水库源水中,以探究其对实际源水总氮的去除效果.结果显示:Sxf14能以硝酸盐和亚硝酸盐为唯一氮源进行好氧反硝化.反应48 h后,NO3--N和NO2--N的去除率分别达74.84(±0.86)%和40.52(±1.49)%,TN去除率最高达到65.07(±1.56)%和41.33(±0.98)%;在以NH4Cl为氮源的异养硝化系统内,该菌在48 h内使NH4+-N浓度由3.73(±0.08)mg/L降到1.28(±0.20)mg/L,氨氮去除率达到65.63(±1.39)%.72 h内,微污染水库源水的TN浓度由2.46(±0.02)mg/L降到1.68(±0.01)mg/L,去除率达到31.7(±0.14)%.因此,菌株Acinetobacter sp.Sxf14具有反硝化能力,能承受较低的碳氮比,降低微污染源水中的氮素,本研究可为微污染水体的菌剂修复技术提供科学依据.     

COD与DO对好氧颗粒污泥同步硝化反硝化脱氮的影响

COD和DO浓度对好氧颗粒污泥的同步硝化反硝化反应有明显影响．COD浓度在400～1200mg／L范围内，好氧颗粒污泥去除COD的能力均在85％以上．颗粒污泥能吸附有机物，使废水中COD浓度快速下降．COD浓度小于800mg／L，好氧颗粒污泥具有良好的脱氮能力，氮去除率最高达85．3％．在溶氧浓度为1-4mg／L条件下，颗粒污泥对COD去除率均在90％以上．不同的溶氧浓度对氮的去除率有一定影响，在溶氧浓度3mg／L时，氮去除率最高，达83％．图7参7     

异养硝化细菌处理氨氮废水及影响因素研究

从生物陶粒反应器中筛选出6株异养硝化细菌，将异养硝化细菌扩大培养后，建立SBR反应器并进行了氨氮去除的试验研究。在SBR反应器进入稳定运行阶段时，可以观察到系统对于氨氮的去除率稳定在82．96％左右，表现出较好的氨氮去除效果；出水亚硝酸盐含量一直维持在较低的水平，其最大值不超过3．84mg·L-1；COD的平均去除率为54．72％，基本实现了同一反应器中的有机物和氨氮的共同去除。异养硝化SBR反应器温度为29℃时，反应器对氨氮和总氮的去除能力最大为82．28％和47．27％；在pH值为8．0时，氨氮去除率最高达到80．15％。C／N〈4．5时，随着C／N比的增加，氨氮和总氮的去除率快速增加；在C/N为6时，氨氮去除率最高达到87．62％。     

一株新的反硝化短程除硫菌的鉴定及主要培养因素筛选

依据反硝化除硫原理,以味精废水污泥为种泥,利用全混流反应器富集并分离出同步反硝化短程除硫菌(SNBI),采用传统与现代分子生物学相结合的手段对其鉴定,以确定其分类地位;同时对SNB1的主要培养因素(营养和环境)进行筛选.结果表明:SNB1的形态特征及生理生化指标与Thauera selenatis最相似,同源性达99.0%,属短杆菌属,尚无中文命名;生理生化指标、富集条件及富集过程物料平衡显示SNB1是一株兼性厌氧反硝化除硫菌;培养SNB1的最佳碳源为蔗糖,最佳氮源为蛋白胨,最佳培养温度为35℃,最适宜pH范围为7～9;最佳条件培养时,OD_(650)和对数细菌数量(CFU)呈直线相关,相关系数R~2=0.981.     

Influence of carbon source and temperature on the denitrifying phosphorus removal process

To supply the valuable operating parameters for the popular usage of the new denitrifying phosphors removal process, it is essential to study the dominant biochemical reactions and the characteristics of denitrifying phosphorus removing bacteria (DPB). Thus, parallel batch experiments using DPB sludge were carried out to assess the effect of substrates (sewage, HAc, and endogenous carbon source) on denitrifying dephosphorus removal efficiency in this study. The results showed that the initial specific phosphorus release rate increased with the high concentration of the short-chain volatile fatty acids ratio in the influent, and sufficient phosphorus was released by DPB. This improved the subsequent denitrification and phosphorus uptake efficiency. The specific endogenous denitrification mainly relies on the internal carbon source (PHB) stored by poly-P bacteria. Denitrifying phosphorus removing bacteria were very hungry when the internal PHB was consumed. Consequently, the specific endogenous denitrification rate was low and the phosphorus uptake did not happen. On the other hand, in the experiment, the denitrifying phosphorus removal performance under two temperature conditions (8–10°C and 25–26°C) was also investigated and analyzed. It was found that the lower temperature decreased the specific phosphorus release and uptake rate, but did not inhibit the denitrifying phosphorus removal completely. Therefore, the negative influence of the low temperature on the overall phosphorus removal was not significant.     

Nitrogen removal performance of municipal reverse osmosis concentrate with low C/N ratio by membrane-aerated biofilm reactor

A membrane-aerated biofilm reactor was employed to investigate the nitrogen removal of one typical municipal reverse osmosis(RO) concentrate with a high total nitrogen (TN) concentration and a low C/ N ratio. The effects of operational conditions, including the aeration pressure, the hydraulic retention time and the C/N ratio, on the systematic performance were evaluated. The nitrogen removal mechanism was evaluated by monitoring the effluent concentrations of nitrogen contents. Furthermore, the microbial tolerance with elevated salinity was identified. The results indicated that the optimal TN removal efficiency of 79.2% was achieved of the aeration pressure of 0.02 MPa, hydraulic retention time of 24 h, and the C/N ratio of 5.8, respectively. It is essential to supplement the carbon source for the targeted RO concentrate to promote the denitrification process. The inhibitory effect of salinity on denitrifying bacteria and nitrite oxidizing bacteria was significant, revealing the limited TN removal capacity of the conditions in this work. The TN removal efficiency remained more than 70% with the addition of salt (NaCl) amount below 20 g/L. This work preliminarily demonstrated the MABR feasibility for the nitrogen removal of municipal RO concentrate with low C/N ratio and provided technical guidance for further scale-up application.

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沣河水系脱氮微生物群落结构研究

河流水体氮素的超负荷不仅破坏了水体生态环境,也严重威胁着人类的生存和发展.水体中有机氮、无机氮（氨氮、亚硝氮、硝氮）和分子氮之间的转化（氮循环）有赖于水体中大量的氮循环微生物（固氮细菌、硝化细菌和反硝化细菌）,然而这些氮循环微生物的生长繁殖也受到包括氮素的形态和浓度在内的多种环境因子的影响,这些因素也通过影响氮循环微生物的生长繁殖进而使得水体中氮素的转化速率发生变化,对水体氮污染的防治有不可忽视的作用.本研究通过在沣河设置不同的研究断面,采集水体样品,进行水质分析,并通过现代分子生物学技术（PCR-DGGE）方法对研究断面水体中氮循环微生物（固氮细菌、硝化细菌和反硝化细菌）的群落结构进行分析.再通过统计学软件对所得分子生物学信息与水质环境因子的相关性进行统计学分析,发现沣河水体中氮循环微生物群落结构受到多种环境因子共同影响,且在枯水期和丰水期表现出不同的特征.在丰水期沣河水体中,硝化细菌群落在中游表现出较高的多样性和丰富性,这与沣河中上游农业COD（化学需氧量）、BOD（生化需氧量）氨氮及有机氮污染物排放量较大,沣河水体DO（溶解氧）高有关.水体中的氨氮、亚硝氮、温度的增加是促进水体中硝化细菌的均匀性和丰富度的增高的主要因子,而pH 值的升高,使得水体中硝化细菌的均匀性和丰富度降低.反硝化微生物在中游和下游的多样性和丰富度较高,与有机物及硝酸盐含量相关.水体中的BOD、COD、TP（总磷）、硝氮的增加是促进水体中反硝化细菌的均匀性和丰富度的增高主要相关因子,而DO 的增多则会对部分反硝化细菌产生不利影响,使得水体中反硝化细菌的均匀性和丰富度降低.本研究结果为沣河以及其他河流的污染控制以及基于微生物的生态修复提供了科?     

集约化蔬菜种植区地下水中反硝化细菌的分离鉴定

从研究我国典型集约化蔬菜种植区地下水中硝酸盐的来源和浓度人手,进行富集、培养、分离、纯化,筛选出一株具有反硝化作用的菌株,通过形态学、革兰氏染色结合16SrDNA序列同源性分析鉴定,其鉴定结果为农杆菌(Agrobacterium sp.).该研究为开展地下水硝酸盐污染的生物修复储备宝贵的菌种资源,为地下水中硝酸盐污染的原位微生物修复和相关污水的生物处理提供微牛物基础,对于经济、有效的解决地下水硝酸盐污染和水资源短缺的问题有着十分重要的意义.     

Effects of La3+, Ce3+ on nitrogen removal in sequencing batch reactor

Batch experiments were conducted to study the short-term biological effects of rare earth ions (La³⁺, Ce³⁺) and their mixture on the nitrogen removal in a sequencing batch reactor (SBR). The data showed that higher NH₄ ⁺-N removal rate, total inorganic nitrogen removal efficiency, and denitrification efficiency were achieved at lower concentrations of rare earth elements (REEs) (<1 mg/L). In the first hour of the aeration stage of SBR, the presence of REEs increased the total inorganic nitrogen removal efficiency and NH₄ ⁺-N removal efficiency by 15.7% and 10%–15%, respectively. When the concentrations of REEs were higher than 1 mg/L, the total inorganic nitrogen removal efficiency decreased, and nitrate was found to accumulate in the effluent. When the concentrations of REEs was up to 50.0 mg/L, the total inorganic nitrogen removal efficiency was less than 30% of the control efficiency with a high level of nitrate. Lower concentrations of REEs were found to accelerate the nitrogen conversion and removal in SBR.     

Promotive effect of pyridine on indole degradation by activated sludge under anoxic conditions

Batch experiments were carried out to investigate the promotive effect of pyridine on indole degradation under denitrifying conditions. The seed sludge was obtained from a local coal-coking wastewater treatment facility and was acclimated in the laboratory. Indole and pyridine were supplemented to the synthetic wastewater at different ratios. The optimum ratio of chemical oxygen demand (COD) to nitrate (C/N) was 8.4–8.9 for both denitrification and indole and pyridine degradation. At a temperature of 28°C and pH of 7.0–7.5, the nitrate reductase activity (NRA) was in the best state. The addition of pyridine could promote NRA and the degradation of indole. When the initial concentration of indole was 150 mg/L, the concentration ratio of indole to pyridine was in the range of 1–10. Under optimum C/N conditions, the degradation of indole could be described with pseudo-zero-order kinetics. There was no accumulation of nitrite during the reaction. When the concentration ratio of pyridine to indole was less than 0.25 with an increase in the pyridine proportion, there were more significant augment rates for NRA and the degradation of indole than the situation when the concentration ratio was more than 0.25.