好氧反硝化细菌的筛选及反硝化特性研究

目的：筛选好氧反硝化细菌,减少水体的亚硝态氮污染。方法：通过BTB平板初筛及反硝化培养基复筛得到目的菌,并探讨了不同溶解氧浓度、自然水体环境对该菌株反硝化作用的影响。结果：分离到1株高效的好氧反硝化细菌A1,该菌的反硝化作用主要发生在菌体的对数生长期,在溶解氧浓度为5mg/L时,对亚硝酸盐氮的降解率达到99％,在自然水体环境中当碳氮摩尔比为10：1时,对亚硝酸盐氮降解率达99％。结论：筛选到一株高效好氧反硝化细菌A1,将其应用于治理养殖水体的亚硝态氮污染有广阔的前景。     

耐冷好氧亚硝酸盐型反硝化细菌的鉴定及脱氮特性研究

旨在对获得的具有耐冷高效去除亚硝酸盐氮和总氮的好氧反硝化细菌进行分类地位及除氮特性研究。结合形态学观察、特异性磷脂脂肪酸检测和16S r RNA基因序列分析,对实验室新分离获得的Y-9菌株进行鉴定,在此基础上,研究了温度、转速、p H、接种量、碳源和亚硝酸盐氮浓度等不同条件对该菌脱氮能力的影响。结果显示,菌株Y-9属于恶臭假单胞菌(Pseudomonas putida),还原亚硝酸盐氮的最适温度为15℃,最佳溶解氧水平对应转速为100 r/min,在该溶解氧水平下15℃、48 h内对亚硝酸盐氮和总氮的去除率高达100%和77.13%,最佳p H为7,100 m L反硝化培养基中最适接种量为1 m L OD600为0.5的菌悬液,碳源为柠檬酸钠,适合治理低浓度的亚硝酸盐氮污染水体,但对高浓度的亚硝酸盐氮具有一定耐受性。Y-9为一株耐冷反硝化细菌,在亚硝酸盐污染的养殖水体,尤其是冷水鱼类养殖水体中具有较大的应用潜力。     

海洋着色菌YL28对含沉积物的海水养殖水体氮污染去除效应的研究

用含有沉积物的氮化物污染实际水体作为测定体系,通过测定水体无机三态氮、总氮、COD、pH以及沉积物总氮、COD和对4种代谢酶活性的动力学过程的监测,评价了一株不产氧光合细菌——海洋着色菌(Marichromatium gracile)YL28对海水实际养殖水体脱氮效应。结果表明:水体本身具有一定的自净作用;与对照组相比,YL28组水体氨氮、亚硝氮、硝氮、沉积物总氮和COD的残留率明显降低(P0.05),在6~10d内水体COD明显升高,水体总氮和pH未见显著差异。处理过程中,在特定的时间范围内,转化酶、蛋白酶、脲酶和脱氢酶活性明显升高(P0.05),与沉积物总氮和COD去除规律相吻合。由此可见,添加YL28对污染海水养殖水体中无机三态氮、沉积物有机氮化物和有机碳化物具有明显的去除作用,本研究为海水养殖微生态水质修复剂的开发和应用奠定基础。     

缢蛏养殖池塘氨氧化微生物的季节变化特征

为探究缢蛏(Sinonovacula constricta)养殖池塘中氨氧化细菌(AOB)和氨氧化古菌(AOA)的季节变化及其环境因子相关性, 试验以amoA基因为分子标记, 利用荧光定量PCR技术对养殖池塘水体及沉积物中的AOA和AOB进行了研究。结果显示, 季节变化对于水体及沉积物中的AOA和AOB的amoA基因丰度有一定的影响, 不同季节的水体及沉积物中AOA含量均比AOB含量高一个数量级。其中水体中AOB在秋季显著高于其余三季(P<0.05); AOA在冬季最低, 且显著低于春秋两季(P<0.05)。沉积物中AOB和AOA均在冬季显著高于其余三季(P<0.05)。环境因子相关性分析结果表明: 水体中AOB丰度与氨氮成显著正相关(P<0.05), 与亚硝氮成显著负相关(P<0.05); AOA丰度与水温成正显著相关(P<0.05), 与硝氮成负显著相关(P<0.05)。沉积物中AOB丰度与溶解氧、硝氮、总氮及亚硝氮成显著正相关(P<0.05), 与水温成显著负相关(P<0.05); AOA丰度与溶解氧、总氮、硝氮成显著性正相关(P<0.05), 与水温成负显著相关(P<0.05)。多元线性回归方程结果显示, 缢蛏养殖池塘的AOA及AOB丰度与硝氮密切相关。研究表明, AOA在缢蛏养殖池塘氨氧化进程中占据主导地位, AOA和AOB丰度受盐度、温度、氮素浓度及溶解氧等环境因子的共同影响。     

好氧反硝化细菌的鉴定及其脱氮特性研究

2018年3月,在广东省中山市的草鱼(Ctenopharyngodon idellus)养殖池塘水体中分离筛选出一株高效好氧反硝化细菌,结合菌株的形态观察、生理生化特性和16S rDNA基因序列分析鉴定为Pseudomonas furukawaii,命名为ZS1。并进一步研究了该菌株的脱氮特性,同时采用单因素实验方法探究了不同碳源种类、温度、pH、C/N和摇床转速对菌株ZS1脱氮效率的影响。结果表明, ZS1菌株在好氧条件下具有高效的脱氮效-N)的含量从48.93降低到1.27 mg/L,去除率为97.40%,去除速率达0.993mg/(L·h);总氮(TN)的含量从52.04降低到8.40 mg/L,去除率为83.86%,去除速率达0.909 mg/(L·h),且无亚硝酸-N为唯一氮源,菌株ZS1发挥最佳好氧反硝化性能的碳源为乙酸钠、柠檬酸钠和葡萄糖、温度为25—35℃、pH为7.0—10.0、C/N为15—25、转速为100—200 r/min。上述结果显示,菌株P.furukawaii ZS1具有良好的好氧反硝化性能,将为池塘养殖尾水处理应用生物脱氮技术提供理论依据及成为初步的候选菌株。     

海河流域水体沉积物碳、氮、磷分布与污染评价

从全流域尺度上研究海河流域水体沉积物碳、氮、磷元素含量与分布特征,对研究海河流域水环境污染现状具有重要意义.本研究采集海河流域河流与水库163个表层沉积物(0～10 cm)样品,测定沉积物有机碳(TOC)、总氮(TN)、总磷(TP)的含量,运用有机指数与有机氮方法评价沉积物污染状况.结果表明: 海河流域沉积物TOC、TN、TP含量具有较大的空间异质性,下游平原区明显高于山区,北四河下游平原、黑龙港及运东平原含量较高,永定河山区、北三河山区、滦河流域含量相对较低,河流沉积物TOC含量显著高于水库沉积物,而TN、TP含量与水库沉积物没有显著差异.TN与TOC、TP含量呈显著正相关(r=0.704,P<0.01;r=0.250,P<0.01).全流域有机指数总体属于“较清洁”水平,北四河下游平原总体已接近有机污染水平.全流域有机氮总体处在“尚清洁”水平,北四河下游平原、黑龙港及运东平原流域存在有机氮污染.海河流域河流、水库沉积物具有相似的污染强度.沉积物C/N平均值为12.71,表明TOC多来源于藻类等浮游动植物,其次是高等植物,水库C/N值比河流高,陆源物质输入对水库沉积物TOC的贡献比对河流大.     

高效净水异养硝化细菌的筛选鉴定及固定化应用研究

本研究从海参养殖水体、泥土中筛选出4株具有硝化能力的异养硝化细菌。分别将其游离菌体细胞投入海参养殖水体,测定亚硝态氮、氨氮去除率,筛选出HS．NOB2为高效净化菌株,对HS-NOB2进行16SrDNA扩增及序列测定,初步鉴定为节杆菌（Arthrobactersp．）。利用海藻酸钠包埋法对高效净化菌体细胞进行固定化,将该固定化菌投入养殖水体及人工合成污水,研究其对水体中亚硝态氮、氨氮的处理效果,并与游离菌体细胞进行比较。结果表明,固定化后亚硝态氮去除率达到49．85％,氨氮去除率达到56．58％,均明显高于游离菌体细胞。上述研究为探寻水体净化提供了新思路,为水质改良剂的实际生产提供可选菌株。     

4种理化因子对菌株XH1硝化效果的影响

【背景】基于硝化菌群的富集培养技术可高效稳定地去除养殖水体中的有害氮素,而当前在水产养殖领域有关硝化菌群定向培育及硝化功能菌株的研究较少。【目的】研究不同盐度、pH、温度、通气量条件下硝化菌群分离菌株XH1的生长及其对氨氮和亚硝氮的去除效果。【方法】设置不同梯度的盐度、pH、温度、通气量条件,通过计数菌量、测定氨氮及亚硝氮的浓度变化,比较不同条件下菌株XH1的生长及其对氨氮和亚硝氮的影响。【结果】菌株XH1可在盐度5‰-35‰、pH 6.0-9.0、温度15-45°C和通气量0.5-1 V/(V·min)的条件下生长良好,菌量最高可达2.34×109cells/mL;在盐度5‰-35‰、pH 6.0-9.0、温度15-30°C、通气量0.5 V/(V·min)的条件下,对氨氮的去除效果显著(P0.05),在第1-3天对培养液中氨氮的最高去除率可达86%-97%,但培养液中的氨氮浓度先降后升;对亚硝氮的最高去除率达68%。【结论】菌株XH1对盐度、pH、温度等主要环境因子具有良好的适应性,其对水体氨氮的去除效果良好,可作为中低盐度养殖池塘水体氨氮防控菌剂产品研发的备选菌株。     

反硝化除磷菌筛选及其特性研究

【目的】研究反硝化除磷菌特性。【方法】通过微生物筛选和生物学特性研究方法,从对虾养殖池塘中筛选出多株可在有氧条件下同时具有反硝化除磷功能的菌种。【结果】菌株LY-1可在18 h内将初始量为10 mg/L的亚硝酸盐氮降低至0.04 mg/L,PO43?-P降低至0.05 mg/L。在DO浓度为5.0?5.9 mg/L时,该菌反硝化除磷率近100%。试验选取具有反硝化除磷功能的枯草芽孢杆菌为阳性对照菌,大肠杆菌为阴性对照菌,比较研究了菌株LY-1在不同pH、温度、盐度、PO43?-P浓度、亚硝酸盐浓度时反硝化除磷的强弱,在pH为5?9范围时,该菌亚硝酸盐氮去除率近99%,PO43?-P去除率86%;温度为30°C时,该菌反硝化除磷率近100%;盐度为5‰?15‰、PO43?-P浓度为10 mg/L、亚硝酸盐氮浓度为20 mg/L时,该菌亚硝酸盐氮和PO43?-P去除率均可达99%。【结论】菌株LY-1反硝化除磷性能显著高于对照菌(P<0.05)。通过菌株LY-1形态学观察、生理生化及16S rRNA基因序列分析,初步鉴定为蜡样芽孢杆菌(Bacillus cereus)。     

耐冷亚硝酸盐型反硝化菌Pseudomonas tolaasii Y-11的鉴定及其脱氮特性

摘要:【目的】反硝化细菌在生物脱氮中具有重要作用,而耐冷亚硝酸盐型反硝化细菌研究较少,本文从长期淹水的冬水田泥土分离获得一株耐冷高效去除亚硝酸盐氮和总氮的好氧反硝化细菌Y-11,明确其分类地位以及除氮特性,以期为后续利用该菌在初冬到春末处理亚硝酸盐水体污染奠定基础。【方法】通过形态学特征、特异性磷脂脂肪酸以及16S rRNA基因测序分析对该菌株进行鉴定;在好氧条件下以亚硝酸钠为唯一氮源,分别研究不同初始温度、转速、pH、碳源、接种量以及亚硝酸盐氮浓度对该菌去除亚硝酸盐氮和总氮的影响,确定最适降解条件。【结果】分离得到的菌株Y-11,经鉴定归于托拉斯假单胞菌(Pseudomonas tolaasii);在国内外尚无该种菌具有反硝化作用的报道,是对亚硝酸盐型反硝化细菌的进一步补充。Y-11菌株的最适脱氮条件为15 ℃,200 r/min,pH7.0,100 mL反硝化培养基中最适接种量为1.5×108 CFU,最佳碳源为乙酸 钠,亚硝酸盐氮为10 mg/L;以乙酸钠为电子供体,15 ℃、初始pH为7.2、150 r/min 振荡培养,48 h对亚硝酸盐氮和总氮的去除率分别为100%和61.28%。【结论】Y-11是一株具有较高反硝化能力的托拉斯假单胞菌,能高效地去除亚硝酸盐氮和总氮,其最适温度是15 ℃左右,是一株耐冷反硝化细菌。     

Sequencing batch membrane biofilm reactor for simultaneous nitrogen and phosphorus removal: novel application of membrane-aerated biofilm

A sequencing batch membrane biofilm reactor (SBMBfR) was developed for simultaneous carbon, nitrogen, and phosphorus removal from wastewater. This reactor was composed of two functional parts: (1) a gas-permeable membrane on which a nitrifying biofilm formed and (2) a bulk solution in which bacteria, mainly denitrifying polyphosphate-accumulating organisms (DNPAOs), were suspended. The reactor was operated sequentially under anaerobic condition and then under membrane aeration condition in one cycle. During the anaerobic period, organic carbon was consumed by DNPAOs; this was accompanied by phosphate release. During the subsequent membrane aeration period, nitrifying bacteria utilized oxygen supplied directly to them from the inside of the membrane. Consequently, the nitrite and nitrate products diffused into the bulk solution, where they were used by DNPAOs as electron acceptors for phosphate uptake. In a long-term sequencing batch operation, the mean removal efficiencies of total organic carbon (TOC), total nitrogen (T-N), and total phosphorus (T-P) under steady-state condition were 99%, 96%, and 90%, respectively. In addition, fluorescence in situ hybridization (FISH) clearly demonstrated the difference in bacterial community structure between the membrane biofilm and the suspended sludge: ammonia-oxidizing bacteria belonging to the Nitrosomonas group were dominant in the region adjacent to the membrane throughout the operation, and the occupation ratio of the well-known polyphosphate-accumulating organism (PAO) Candidatus "Accumulibacter phosphates" in the suspended sludge gradually increased to a maximum of 37%.     

池塘氮循环中各种细菌与理化因子的相关性研究

对精养鱼池的水体及淤泥进行多次采样,用统计学分析处理,结果表明：硝化类细菌总体上与各主要理化因子的相关性较强（R＞0．60）,其中氨化菌主要与溶氧相关程度较高;反硝化菌主要与有效磷相关程度较高;亚硝化菌主要与氨氮负相关程度较高;硝化菌主要与亚硝酸盐相关程度较高。硝化类细菌与异养菌之间的相关性则更强,如反硝化菌与厌养菌（R＝0．944,P＝0．001）、拓硝化菌和好氧异养菌（R＝0．832,P＝0．003）皆显正相关,亚硝化菌和厌氧异养菌（R＝－0．76,P＝0．009）显负相关;而在硝化类细菌之间的相关程度却较弱（R＜0．60）,表明池塘硝化类细菌对水质具有一定的调控作用,但相互之间的依赖性不强,各自相对独立地发挥作用。     

基于好氧反硝化及反硝化聚磷菌强化的低温低碳氮比生活污水生物处理中试研究

【背景】低碳氮比生活污水很难达标处理,多级A/O工艺、生物强化技术及生物膜技术的有机结合可有效解决这一问题。【目的】开发出一种泥膜共生多级A/O工艺并进行中试研究,驯化出高效脱氮除磷菌剂并对系统进行生物强化。【方法】通过测定中试设备出水及污水处理厂出水化学需氧量(Chemical oxygen demand,COD)、氨氮(NH_4~+-N)、硝氮(NO_3~--N)、总氮(Total nitrogen,TN)、总磷(Total phosphorus,TP)对比分析两种工艺的污染物去除效能,利用高通量测序技术对比生物强化技术对系统微生物群落结构的影响。【结果】中试设备对COD、NH_4~+-N、NO_3~--N、TN、TP的去除效果均优于污水处理厂的处理工艺;驯化的低温好氧反硝化菌TN去除率最大值可达84.21%,驯化的低温反硝化聚磷菌群对磷的去除率最高可达85.75%;利用驯化菌群对中试设备进行生物强化后较好地改善了系统NH_4~+-N、NO_3~--N、TN、TP的去除效果;经生物强化后,具有好氧反硝化和反硝化聚磷功能的Pseudomonas菌群明显增多。【结论】泥膜共生多级A/O工艺对于低碳氮比生活污水的处理具有很好的效果,利用生物强化技术可有效提高低温条件下系统污染物去除效能。     

Nitrifying bacterial growth inhibition in the presence of algae and cyanobacteria

Nitrifying bacteria, cyanobacteria, and algae are important microorganisms in open pond wastewater treatment systems. Nitrification involving the sequential oxidation of ammonia to nitrite and nitrate, mainly due to autotrophic nitrifying bacteria, is essential to biological nitrogen removal in wastewater and global nitrogen cycling. A continuous flow autotrophic bioreactor was initially designed for nitrifying bacterial growth only. In the presence of cyanobacteria and algae, we monitored both the microbial activity by measuring specific oxygen production rate (SOPR) for microalgae and cyanobacteria and specific oxygen uptake rate (SOUR) for nitrifying bacteria. The growth of cyanobacteria and algae inhibited the maximum nitrification rate by a factor of 4 although the ammonium nitrogen fed to the reactor was almost completely removed. Terminal restriction fragment length polymorphism (T‐RFLP) analysis indicated that the community structures of nitrifying bacteria remained unchanged, containing the dominant Nitrosospira, Nitrospira, and Nitrobacter species. PCR amplification coupled with cloning and sequencing analysis resulted in identifying Chlorella emersonii and an uncultured cyanobacterium as the dominant species in the autotrophic bioreactor. Notwithstanding their fast growth rate and their toxicity to nitrifiers, microalgae and cyanobacteria were more easily lost in effluent than nitrifying bacteria because of their poor settling characteristics. The microorganisms were able to grow together in the bioreactor with constant individual biomass fractions because of the uncoupled solids retention times for algae/cyanobacteria and nitrifiers. The results indicate that compared to conventional wastewater treatment systems, longer solids retention times (e.g., by a factor of 4) should be considered in phototrophic bioreactors for complete nitrification and nitrogen removal. Biotechnol. Bioeng. 2010;107: 1004–1011. © 2010 Wiley Periodicals, Inc.     

Characteristics and mechanisms of the hydroponic bio-filter method for purification of eutrophic surface water

The hydroponic bio-filter method (HBFM) was adopted to purify eutrophic surface water. The average removal efficiency of total nitrogen (TN) and total phosphorus (TP) was 16.8% and 30.8%, respectively, at the hydraulic loading rate (HLR) of 3.0 m³ (m² d)⁻¹. The mass removal rate of TN and TP accordingly reached 1.0 and 0.1 g (m² d)⁻¹ separately. The sedimentation of particulate nitrogen and phosphorus played a major role in removal of nitrogen and phosphorus, which contribute 62.2% and 75.9%, respectively. The optimal HLR of HBFM ranged from 3.0 to 4.0 m³ (m² d)⁻¹. The sediment in midstream reached a maximum nitrification potential of 4.76 × 10⁻⁶ g (g h)⁻¹, while upstream it reached a maximum denitrification potential of 8.1 × 10⁻⁷ g (g h)⁻¹. The distribution of nitrification potential corresponded to the ammonium-oxidizing bacteria density. The key for improving nitrogen removal efficacy of HBFM system consisted of changing the nitrification/denitrification region distribution and accordingly enhancing the denitrification process. The sum of dissolved nitrogen removed by denitrification and plant assimilation was nearly equal to the amount released by sediment. Secateur length of Nasturtium officinale had some effect on its uptake rate. The length of cut should be less than 10 cm at a time. The harvesting frequency of once a month for N. officinale had no influence on nitrogen and phosphorus removal.     

Denitrifying phosphorus removal and impact of nitrite accumulation on phosphorus removal in a continuous anaerobic-anoxic-aerobic (A2O) process treating domestic wastewater

A lab-scale anaerobic-anoxic-aerobic (A(2)O) process was operated to investigate denitrifying phosphorus removal and nitritation-denitritation from domestic wastewater, especially regarding the impact of nitrite accumulation caused by nitritation on phosphorus removal. The results showed that mean total nitrogen (TN) removal was only about 47% and phosphorus removal was almost zero without the pre-anoxic zone and additional carbon source. Contrastively, with configuration of pre-anoxic zone, TN and phosphorus removal was increased to 75% and 98%, respectively, as well as denitrifying phosphorus removal of 66-91% occurred in the anoxic zone. Nitritation-denitritation was achieved through a combination of short aerobic actual hydraulic retention time and low dissolved oxygen levels (0.3-0.5 mg/L); however, phosphorus removal deteriorated with increase of nitrite accumulation rates. The free nitrous acid (FNA) concentration of 0.002-0.003 mg HNO(2)-N/L in the aerobic zone inhibited phosphorus uptake, which was major cause of phosphorus removal deterioration. Through supplying the carbon sources to enhance denitrification and anaerobic phosphorus release, nitrite and FNA concentrations in the aerobic zone were reduced, and phosphorus removal was improved. Compared with nitrification-denitrification, nitritation-denitritation reduced the carbon requirement by 30% and performed biological nutrients removal well with mean TN and phosphorus removal of 85% and 96%, respectively.     

Anammox enrichment from reject water on blank biofilm carriers and carriers containing nitrifying biomass: operation of two moving bed biofilm reactors (MBBR)

The anammox bacteria were enriched from reject water of anaerobic digestion of municipal wastewater sludge onto moving bed biofilm reactor (MBBR) system carriers-the ones initially containing no biomass (MBBR1) as well as the ones containing nitrifying biomass (MBBR2). Duration of start-up periods of the both reactors was similar (about 100?days), but stable total nitrogen (TN) removal efficiency occurred earlier in the system containing nitrifying biomass. Anammox TN removal efficiency of 70% was achieved by 180?days in both 20?l volume reactors at moderate temperature of 26.0°C. During the steady state phase of operation of MBBRs the average TN removal efficiencies and maximum TN removal rates in MBBR1 were 80% (1,000?g-N/m(3)/day, achieved by 308?days) and in MBBR2 85% (1,100?g-N/m(3)/day, achieved by 266?days). In both reactors mixed bacterial cultures were detected. Uncultured Planctomycetales bacterium clone P4, Candidatus Nitrospira defluvii and uncultured Nitrospira sp. clone 53 were identified by PCR-DGGE from the system initially containing blank biofilm carriers as well as from the nitrifying biofilm system; from the latter in addition to these also uncultured ammonium oxidizing bacterium clone W1 and Nitrospira sp. clone S1-62 were detected. FISH analysis revealed that anammox microorganisms were located in clusters in the biofilm. Using previously grown nitrifying biofilm matrix for anammox enrichment has some benefits over starting up the process from zero, such as less time for enrichment and protection against severe inhibitions in case of high substrate loading rates.     

进水碳氮比对CANON型人工湿地脱氮性能的影响

通过逐步提高进水中的有机碳源浓度,探讨进水碳氮比(C/N)对基于亚硝化的全程自养脱氮(CANON)型潮汐流人工湿地(TFCW)脱氮效能及其微生物特性的影响.结果表明: 进水C/N可显著影响CANON型TFCW中脱氮功能微生物的数量与活性,进而影响其氮素转化速率.当进水C/N由0.0增至6.0时,TFCW中反硝化功能基因的丰度随之增加,系统反硝化性能提高,TFCW中逐渐形成同步亚硝化、厌氧氨氧化与反硝化(SNAD)耦合反应体系,其脱氮效果得以强化.当进水C/N＞6.0时,好氧氨氧化菌活性受到抑制,数量逐渐减少,TFCW中的厌氧氨氧化作用与反硝化作用受阻,系统脱氮性能恶化.当进水C/N为6.0时,TFCW中的SNAD作用可得到最大限度的强化,其总氮(TN)去除率和去除负荷分别达(93.3±2.3)%和(149.30±8.00) mg·L^-1·d^-1,高于CANON系统中TN去除率的理论值.