好氧反硝化菌

<正>养殖水体氮素污染问题是目前困扰我国水产养殖业可持续发展的一大难题。生物脱氮技术被认为是目前最具发展前景的水体脱氮技术,其效果的优劣与所采用菌株的特性密切相关。传统反硝化细菌仅能在厌氧及低氧条件下发挥脱氮作用,与养殖水体的高溶氧环境矛盾,而好氧反硝化细菌则可在高溶氧环境中发挥脱氮作用,显著提高生物脱氮技术在养殖水体中的应用效果,实现养殖水体的绿色、零污染脱氮。因而,对好氧反硝化细菌开展高效选育方法的研究,找到可适应养殖水体水环境的微生物菌株具有重要的理论价值和经济价值。     

耐高浓度氨氮的异养硝化好氧反硝化菌株U1的鉴定及其脱氮特性

【背景】城市垃圾渗滤液是一种成分复杂的有机废水,含氮量高,如果未经处理直接排放到环境中会造成严重的环境污染。【目的】筛选可以耐受垃圾渗滤液中高浓度氨氮并高效去除污水中氮素的异养硝化好氧反硝化菌株,为解决垃圾渗滤液的氮素污染提供功能菌株。【方法】从垃圾渗滤液中筛选分离能耐受高氨氮浓度的菌株,通过测定各菌株的脱氮能力,筛选到一株脱氮能力最强的菌株,命名为U1,通过测定16S rRNA基因序列和生理生化特性确定该菌株为铜绿假单胞菌。进一步研究了菌株U1在不同初始氨氮浓度、碳源、转速、初始pH、碳氮比等单因素变量下的脱氮能力,并结合L₉（3⁴）正交试验研究了菌株U1的最佳脱氮条件。【结果】分离出一株铜绿假单胞菌并命名为U1。该菌株的最优脱氮条件为:初始氨氮浓度为1 000 mg/L,红糖和柠檬酸三钠的混合碳源,pH 6.0,C/N为10,转速为130 r/min,菌株U1的最大总氮去除率为64.37%,最大氨氮去除率为76.73%。对于总氮和氨氮含量分别是2 345 mg/L和1 473.8 mg/L的垃圾渗滤液,菌株U1最大总氮去除率为27.86%...     

一株安全高效的好氧反硝化菌Pseudomonas stutzeri DZ11的生物安全性及脱氮性能研究

对中山市民众镇水产养殖场底泥及污水中分离筛选出的好氧反硝化细菌进行研究,旨在为开发养殖废水的脱氮工艺奠定基础。从样品中筛选出高性能菌DZ11,对其进行形态学观察,生理生化测定以及16S rDNA基因序列分析,确定菌株的种属;进行药敏试验和生物安全性检测,确认菌株的安全性;从碳源种类、温度、pH值和碳氮比这4个因素对脱氮效率的影响进行研究,提高菌株的脱氮效率。鉴定结果表明,DZ11为假单胞菌属施氏假单胞菌,并命名为Pseudomonas stutzeri DZ11。经药敏试验和生物安全性检测发现该菌株无明显耐药性并具有较高的生物安全性。单因素优化的结果为：菌株DZ11的最佳碳源为柠檬酸钠,最适温度35.0℃,最佳pH为7,最佳的C/N为10。在分别以氨氮,硝态氮和亚硝态氮为唯一底物时的转化率最高可分别达到84.39%,99.0%和97.65%,说明菌株具有良好的脱氮作用。菌株DZ11具有较高的生物安全性和高效的脱氮性能,在养殖废水的脱氮工艺中具有较大的应用潜力。     

一株海洋好氧反硝化细菌的鉴定及其好氧反硝化特性

【目的】从处理海洋养殖循环水的生物滤器生物膜中分离到1株具有好氧反硝化活性的细菌(菌株2-8),并进一步研究了该菌的分类地位及反硝化特性。【方法】采用16S rRNA基因序列分析对菌株进行初步鉴定,采用好氧培养技术,探讨了碳源种类、起始pH、NaCl浓度、C/N、温度和摇床转速对菌株2-8好氧反硝化活性的影响。【结果】该菌株的16S rRNA基因序列与Pseudomonas segetis FR1439T(AY770691)的相似性最高,达到99.9%,因此初步鉴定菌株2-8属于假单胞菌属(Pseudomonas sp.2-8)。碳源类型和C/N对其好氧反硝化作用的影响最为显著,以柠檬酸钠为唯一碳源,C/N为15时脱氮效率最高,低C/N导致亚硝酸盐的积累;其好氧反硝化的最适温度和pH分别为30℃和7.5;菌株2-8在摇床转速为160r/min下脱氮效果最好;NaCl浓度对其反硝化活性的影响不明显。【结论】在初始硝酸氮浓度为140mg/L,以柠檬酸钠为唯一碳源、C/N为15、pH为7.5、NaCl浓度为30g/L,30℃以及160r/min摇床培养的条件下,菌株2-8在48h内脱氮率可达92%且无亚硝酸盐积累。     

生物滤塔体系好氧反硝化菌的分离鉴定与特性研究

从实验室生物滤塔填料的生物膜上,经选择性培养基筛选,分离出4株好氧反硝化茵.好氧状态下4种菌的40h反硝化率均大于80%,其中菌种A1反硝化率可达到99.05%.跟踪菌种反硝化过程中氮元素24h变化过程,发现4株菌除A1外都有亚硝酸根积累.茵种A1为短杆菌,革兰氏阴性.生理生化特性研究与16S rDNA序列测定(GenBank接受号DQ836052.1)初步判定菌种A1为假单胞菌Pseudomonas putida.适于茵Al生长的初始pH值是7.0左右,温度30℃左右,当DO大于2.0mg/L时,DO的变化对菌种Al的反硝化效果影响很小.     

海洋环境异养硝化-好氧反硝化菌的研究进展

异养硝化-好氧反硝化（heterotrophic nitrifying-aerobic denitrification,HN-AD）菌的发现打破了传统的脱氮理论,可以在有氧条件下同时进行硝化和反硝化,成为近年来的研究热点。HN-AD细菌在海洋氮循环中发挥着重要作用。本文对海洋环境中HN-AD菌的多样性和部分已知氮代谢途径及相关酶系进行了介绍,分析了盐度、碳氮比、溶解氧、pH等环境因素对HN-AD菌脱氮效果的影响,对其工艺和技术应用、前景和发展方向进行了综述和展望。     

好氧反硝化菌的筛选及其脱氮除磷性质的研究

利用富集培养基, 从用生活污水驯化后的活性污泥中筛选得到一株具有好氧反硝化兼具除磷功能的细菌。通过形态学及生理生化指标鉴定其为假单胞菌属。利用此好氧反硝化菌处理模拟废水及生活废水, 通过监测总氮、无机磷及CODcr变化确定在C/N摩尔比为3:1、接种量为10%、pH 6.8、30°C条件下处理2 d, 该菌株脱氮、除磷及去除有机物的效果最佳, 活性污泥经此好氧反硝化菌强化后, 对生活废水的处理能力得到明显提升。     

生物滤塔体系好氧反硝化菌的分离鉴定与特性研究

从实验室生物滤塔填料的生物膜上, 经选择性培养基筛选, 分离出4株好氧反硝化菌。好氧状态下4种菌的40 h反硝化率均大于80%, 其中菌种A1反硝化率可达到99.05%。跟踪菌种反硝化过程中氮元素24 h变化过程, 发现4株菌除A1外都有亚硝酸根积累。菌种A1为短杆菌, 革兰氏阴性。生理生化特性研究与16S rDNA 序列测定(GenBank接受号DQ836052.1)初步判定菌种A1为假单胞菌Pseudomonas putida。适于菌A1生长的初始pH值是7.0左右, 温度30℃左右, 当DO大于2.0 mg/L时, DO的变化对菌种A1的反硝化效果影响很小。     

分离于河口区芦苇湿地1株好氧反硝化菌的鉴定及其反硝化特性

【目的】筛选高效脱氮且N_2O释放量少的好氧反硝化细菌,并对菌株的反硝化特性进行研究,可为河口湿地富营养化水体的生物修复提供技术支撑。【方法】经BTB培养基初筛和反硝化能力测定,从辽河河口区芦苇湿地土壤中分离得到1株具有较高反硝化能力的好氧反硝化菌C3。经形态观察、生理生化鉴定和16S rRNA序列分析,对菌株进行鉴定。研究温度、碳源、pH及C/N对其生长量、反硝化能力及N2O释放的影响。【结果】筛选得到的高效好氧反硝化细菌C3,经鉴定属于假单胞菌属(Pseudomonas sp.)。反硝化特性研究结果表明,该菌最适碳源为柠檬酸三钠,在温度为30°C、pH为7.0、C/N为10时生长速率和脱氮效率最高且N_2O释放量较少。在此条件下,该菌在36 h内使NO_3~–由179.55 mg/L降至5.08 mg/L,脱氮率高达97.17%。该菌株在整个反硝化过程中中间产物N_2O的最大累积量较低,为0.22 mg/L。【结论】从湿地土壤中分离所得好氧反硝化菌C3为假单胞菌属的1个种(Pseudomonas sp.),该菌株在高效除氮和低N_2O累积方面均具有明显优势,对后续河口湿地富营养化水体治理具有重要意义。     

一株海水异养硝化-好氧反硝化菌系统发育及脱氮特性

【目的】确定一株分离自海水的异养硝化-好氧反硝化菌的系统发育地位并探索其脱氮特性和机理,以期为解释异养硝化-好氧反硝化机理以及改进海水养殖及废水的生物脱氮工艺提供理论依据。【方法】通过形态观察、生理生化实验和16S rRNA基因序列分析,鉴定该菌株;通过测定菌株在不同无机氮源降解测试液中的生长和脱氮效率,分析其异养硝化和好氧反硝化性能。【结果】经鉴定该菌株属于盐单胞菌属(Halomonas);最适生长条件为盐度3%、pH 8.5、温度28℃、碳氮比10:1,在盐度为15%的培养液中仍能生长;可以同时去除氨氮、亚硝酸氮和硝酸氮,24 h时对NH4+-N、NO2--N、和NO3--N的去除率可分别达到98.29%、99.07%、96.48%,3种形态无机氮同时存在时,会优先利用NH4+-N,且总无机氮去除率较单一存在时更高,说明该菌株可实现同步硝化反硝化。【结论】该分离自海水的异养硝化-好氧反硝化菌属于盐单胞菌属(Halomonas),在高盐环境中仍能生长,同时具有高效的异养硝化和好氧反硝化能力,能够独立完成脱氮的全部过程。     

一株好氧反硝化菌的分离及特性研究

从土壤中分离得到一株好氧反硝化细菌CY1, 该菌株在厌氧和好氧条件下均具有反硝化能力。硝酸盐氮初始浓度为137.25 mg/L, 30 h内硝酸盐氮去除率分别为99.98%(厌氧)和60.16%(好氧)。通过形态学特征、生理生化特性及16S rDNA同源性比较对菌株CY1进行鉴定, 初步判断CY1为泛养副球菌(Paracoccus pantotrophus)。     

一株高效好氧反硝化菌的分离及特性

利用富集培养的方法从南昌市郊某养鱼塘采样分离出22株反硝化细菌,其中8株反硝化率较高,从中选择一株效果最好的作为研究对象,命名为HS-N62,对其生长特性进行了深入研究。结果表明:硝酸盐氮初始浓度为140mg/L,菌株HS-N62在12h内对硝酸盐氮的去除率可达96%,而且没有亚硝酸盐氮的积累。该菌最适生长温度范围为30°C-37°C,最适生长pH范围6.0-8.0,最适C/N比为10:1,并能利用多种碳源生长。运用正交试验探讨了该菌株最适的反硝化条件。反硝化菌株HS-N62还具有较好的除磷能力,12h除磷率达到67.7%(初始磷酸盐浓度57mg/L)。通过形态学特性和生理生化分析以及16S rRNA基因序列分析,菌株HS-N62与Pseudomonas sp.亲缘关系最为接近,相似性达99%,初步鉴定该菌为假单胞菌属(Pseudomonas sp.)。     

Survival of the Aerobic Denitrifier Pseudomonas stutzeri Strain TR2 during Co-Culture with Activated Sludge under Denitrifying Conditions

The aerobic denitrifier Pseudomonas stutzeri TR2 (strain TR2) has the potential to reduce nitrous oxide emissions during the wastewater treatment process. In this application, it is important to find the best competitive survival conditions for strain TR2 in complex ecosystems. To that end, we examined co-cultures of strain TR2 with activated sludge via five passage cultures in a medium derived from treated piggery wastewater that contained a high concentration of ammonium. The results are as follows: (i) The medium supported the proliferation of strain TR2 (P. stutzeri strains) under denitrifying conditions. (ii) Nitrite was a better denitrification substrate than nitrate for TR2 survival. (iii) Strain TR2 also demonstrated strong survival even under aerobic conditions. This suggests that strain TR2 is effectively augmented to the wastewater treatment process, aiding in ammonium-nitrogen removal and reducing nitrous oxide production with a partial nitrification technique in which nitrite accumulates.     

关于好氧反硝化菌筛选方法的研究

采用污泥驯化手段富集好氧反硝化细菌,将得到的驯化污泥分离纯化,共得到105株菌。用测TN的方法对所筛菌株进行初筛,得到25株对TN去除率达到50％以上的菌株。用氮元素轨迹跟踪测定法复筛,证实这25株菌都可以在好氧条件下进行硝酸盐呼吸,其中24株菌的反硝化过程为：NO3^-N→NO2^-N→N2,研究中还发现在反硝化过程中硝酸盐和亚硝酸盐不存在明显竞争被利用的作用。同时还提出了可能实现短程同步硝化反硝化以及在反馈作用的调节下,加快硝化反应速度的观点。     

Isolation and Characterization of Coal Solubilizing Aerobic Microorganisms from Salt Range Coal Mines,Pakistan

Microbial solubilization of coal has been considered as a promising technology to convert raw coal into valuable products. In the present study, initially a total of 50 different aerobic bacterial and fungal isolates have been isolated from soil, coal and water samples of Dulmial Coal Mines, Chakwal, Pakistan, but on the basis of solubilization potential, only four isolates were selected for further study. The intensity of biosolubilization was measured by determining the weight loss of the coal pieces, which was observed to be about 25.93% by Pseudomonas sp. AY2, 36.36% by Bacillus sp. AY3 and 50% by Trichoderma sp. AY6, while Phanerochaete sp. AY5 showed maximum coal solubilization potential i.e. 66.67% in 30 days. UV/Vis spectrum revealed an increase in the pattern of absorbance of all treated samples compared to control referring to solubilization. Fourier transform infrared spectroscopy indicated alterations in the structure of treated coal in comparison to control coal suggesting breakdown in the complex structure of coal. The major absorbance bands in infrared spectroscopy for solubilization product were attributed to carbonyl (1,600 cm^?1), hydroxyl (3,450 cm^?1), cyclane (2,925 cm^?1), ether linkage (1,000–1,300 cm^?1), carboxyl (3,300–2,500 cm^?1) and side chains of aromatic ring (1,000–500 cm^?1). The presence of microorganisms and surface erosion of coal residues compared to control samples were observed by scanning electron microscopy, which suggested that isolated microorganisms were able to survive in coal for a longer period of time. Therefore, the present study concluded that microorganisms isolated from coal mines have excellent potential for coal solubilization which is considered as a crucial step in coal methanogenesis allowing them to be used successfully for in situ methane production to meet future energy demands.     

Aerobic batch degradation of phenol using immobilized Pseudomonas putida

Alginate concentrations between 2 and 4% had little effect on the degradation rate of phenol by alginate-immobilized Pseudomonas putida. Ten-degree shifts from 25°C resulted in approximately 30% slower degradation. Maximal degradation rates were favored at pH 5.5–6.0. The response of degradation rate to increased air flow in the bubble column used was almost linear and an optimal higher than 16 vol vol⁻¹ was indicated, although free cells appeared in the reaction medium above 12 vol vol⁻¹. When the initial phenol concentration was raised, degradation rate was not significantly affected until levels higher than 1200 mg ml⁻¹ where performance was markedly reduced. Increasing the ratio of total bead volume to medium volume gave progressively smaller increases in degradation rate. At a medium volume to total bead volume ratio of 5:1, the maximum degradation rate was 250 mg L⁻¹ h⁻¹. Received 24 November 1998/ Accepted in revised form 27 January 1999     

Isolation and characterisation of ethoprophos-degrading bacteria

An enrichment culture technique was used to isolate bacteria responsible for the enhanced biodegradation of ethoprophos in a soil from Northern Greece. Restriction fragment length polymorphism patterns of the 16S rRNA gene, partial 16S rRNA sequence analysis, and sodium dodecylsulfate-polyacrylamide gel electrophoresis total protein profile analysis were used to characterise the isolated bacteria. Two of the three ethoprophos-degrading cultures were pure and both isolates were classified as strains of Pseudomonas putida (epI and epII). The third culture comprised three distinct components, a strain identical to P. putida epI and two strains with 16S rRNA sequence similarity to Enterobacter strains. Isolate epI effectively removed a fresh ethoprophos addition from both fumigated and non-fumigated soil when introduced at high inoculum density, but removed it only from fumigated soil at low inoculum density. Isolates epI and epII degraded cadusafos, isazofos, isofenphos and fenamiphos, but only at a slow rate. This high substrate specificity was attributed to minor (cadusafos), or major (isazofos, isofenphos, fenamiphos) structural differences from ethoprophos. Studies with (14)C-labelled ethoprophos indicated that isolates epI and epII degraded the nematicide by removing the S-propyl moiety.     

一株精氨酸脱亚胺酶产生菌的筛选及鉴定

精氨酸脱亚胺酶由于具有成为精氨酸营养缺陷型肿瘤如肝细胞瘤,黑素瘤治疗药物的潜力而被国内外多个科研机构进行研究。本文报道本研究室于无锡锡惠公园土样中筛选得到一株产精氨酸脱亚胺酶的菌株901,并对其进行了形态,生理生化特征分析及16S rRNA基因分析,该菌被鉴定为恶臭假单胞菌(Pseudomonas putida)。