新型异养氨氧化菌的分离鉴定及氨氧化特性

从青岛海岸采集淤泥．通过富集培养,以硅胶平板点种分离．经纯化得到一株氨氧化菌H9菌株。该菌可以在营养培养基中生长,是一株异养氨氧化菌,经形态、生理生化特性、16SrRNA序列分析等初步鉴定该菌属于假诺卡氏菌属（Pseudonocardia sp．H9）。该菌株具有将氨氧化为硝酸的能力,明显不同于自养型的硝化作用过程需要亚硝化细菌和硝化细菌的密切协作才能把氨氧化成硝酸的特点。此外,该菌可以在无有机碳的培养基或在有机碳C／N达7或更高的培养基中氧化氨至硝酸;在亚硝化培养基与LB的混合培养基中,在LB占15％以下时,LB越高越有利于氨的氧化和菌的生长。该菌株的生长和氧化氨的最适温度为30℃;pH5．0～10．0,最适为8．0;氨离子摩尔浓度1～40mmol／L,最适为8mmol／L;氯化钠质量分数为0％～8％,最适为3．5％。由于该菌株具有以上的特性和优点．在污水的脱氮处理以及养殖海水的氨氮处理中有较高的应用前景,同时也是研究氨的生物氧化机制的理想菌株。     

巢湖低温高效聚磷菌的筛选及其特性

采用低温富集培养及混合平板分离技术,从巢湖底泥中分离筛选得到2株在低温下仍具有较高效能的菌株D3、D6,经过厌氧/好氧交替培养,2种菌株在低温下(8℃)的除磷率均达到80%以上。在低温下,研究了p H、微量元素对2株菌株生长及除磷率的影响,实验结果表明,2株菌都具有广泛的温度适应性,适宜其生长和除磷的p H为中性偏碱,微量元素的缺乏对2株菌株的生长和除磷效果有不同程度的不良影响。染色观察显示,厌氧培养时菌体内聚羟基丁酸(PHB)颗粒明显增多,转为好氧培养后异染颗粒增多,为典型的聚磷菌特征。经鉴定,2株菌均属假单胞菌属。     

一株反硝化细菌的分离鉴定及其反硝化特性简

从处理城镇污水的移动床生物膜反应器中分离获得一株反硝化细菌D₃，并进一步研究该菌株的系统发育地位及反硝化特性。采用16S rDNA序列分析对菌株进行初步鉴定，探讨了基质浓度、起始pH和温度对菌株反硝化活性的影响。根据形态学特征、生理生化特性及16S rDNA序列测定分析，初步鉴定菌株属于寡养单胞菌属。该菌能利用硝酸钠或亚硝酸钠进行反硝化作用，最佳电子受体是硝酸盐氮，反硝化速率最大为19.86 mg/（L·h），最适生长pH为7.36，最适生长温度为33.5℃。菌株D₃在初始硝态氮浓度为140 mg/L，以乙酸钠为惟一碳源，pH为7.36，温度33.5℃的最优生长条件下，培养10 h进入对数生长期，倍增时间为9.9 h，48 h内硝酸盐还原率达95%。     

活性污泥中高效除臭菌株的驯化和筛选

以复合生物滤池作为生物除臭反应器，活性污泥作为菌源，进行氧化S^2-和NH⁺₄的活性驯化，2周后驯化成功，去除率达80%以上。通过平板划线分离，共分离出5株菌，其中细菌、酵母菌平板上各2株，霉菌平板上1株。将分离出的5株菌分别接种至含S^2-、含NO^-₂和含NH⁺₄的3类液体培养基中进行摇床培养2周，通过对菌株氧化能力的实验研究，筛选出2株净化H₂S的高效菌株，2株净化NH₃的高效菌株，能利用恶臭气体作为基质，具有潜在的应用价值。     

3种水培植物根系分泌的有机酸对氮循环菌的影响

通过收集水培吊兰、空心菜和水芹的根系分泌物,采用液相色谱分析了其中有机酸的种类和含量;并研究了根系分泌物中的有机酸对氮循环菌的影响。结果表明3,种植物根系分泌物中的有机酸对氨化细菌和反硝化细菌的生长具有促进作用,对亚硝化细菌和硝化细菌的生长具有抑制作用。     

一株反硝化细菌的分离鉴定及其反硝化特性

从处理城镇污水的移动床生物膜反应器中分离获得一株反硝化细菌D3,并进一步研究该菌株的系统发育地位及反硝化特性。采用16S rDNA序列分析对菌株进行初步鉴定,探讨了基质浓度、起始pH和温度对菌株反硝化活性的影响。根据形态学特征、生理生化特性及16S rDNA序列测定分析,初步鉴定菌株属于寡养单胞菌属。该菌能利用硝酸钠或亚硝酸钠进行反硝化作用,最佳电子受体是硝酸盐氮,反硝化速率最大为19.86 mg/(L·h),最适生长pH为7.36,最适生长温度为33.5℃。菌株D3在初始硝态氮浓度为140 mg/L,以乙酸钠为惟一碳源,pH为7.36,温度33.5℃的最优生长条件下,培养10 h进入对数生长期,倍增时间为9.9 h,48 h内硝酸盐还原率达95%。     

贫营养异养硝化细菌的分离鉴定及硝化性能研究

从水库底泥中分离出1株贫营养异养硝化细菌,命名为菌株Y11。经菌株生理特性研究和16SrDNA测序,鉴定出菌株Y11属于假单胞菌属细菌(Pseudomonas sp.)。考察了温度、pH及C/N对菌株Y11的硝化性能影响,结果表明,菌株Y11进行硝化作用的最适温度为30℃,最适pH为中性,最适C/N为6.0,在该条件下进行4d的硝化反应,菌株Y11对氨氮去除率可达91.42%。     

膜生物反应器好氧反硝化菌的筛选及鉴定

从某膜生物反应器(MBR)污泥中筛选出反硝化活性较高的好氧反硝化细菌,进行了反硝化活性检测、菌种鉴定和作用机制探索。结果表明,共分离出6株好氧反硝化细菌,在较低的菌浓度(1×105个/mL)、DO为4.2~5.5mg/L的条件下启动脱氮反应,菌株F2、F4、F5在24、48h的总氮去除率分别超过40.29%、67.19%,硝酸盐氮去除率分别在64.21%、83.31%以上;经16SrDNA序列测序和比对,菌株F2、F4、F5分别与苍白杆菌属(Ochrobactrumsp.)、蜡状芽孢杆菌(Bacillus cereus)、布鲁菌属(Brucellasp.)的同源性最高;PCR扩增结果表明,菌株F2、F4、F5中均具有周质硝酸盐还原酶,这几种细菌很可能通过这种酶来实现好氧反硝化。     

一株高效脱硫脱氨氮菌的分离、鉴定及系统发育分析

从运行稳定的能同步脱臭的曝气生物滤池中采集样品，富集分离获得一株能高效脱硫脱氨氮的菌株TS-1。对分离菌株进行形态观察、生理生化试验及16S rRNA基因序列分析，结果表明，该菌株TS-1为革兰氏阳性菌，杆状；菌落在营养琼脂培养基上呈圆形，表面光滑，乳白色半透明；V-P试验阴性，能水解淀粉和明胶，利用柠檬酸盐生长；对菌株进行16S rRNA的PCR扩增，扩增产物测序结果表明分离菌株TS-1与巨大芽孢杆菌（Bacillus megaterium）同源性达到99%；以16S rRNA同源性为基础构建了包括24株相关种属细菌在内的系统发育树，在系统发育树中，分离菌株TS1与Bacillus megaterium在同一分支。结合形态观察、生理生化试验及16S rRNA基因序列分析结果，将其初步鉴定为巨大芽孢杆菌（Bacillus megaterium）。在常温（30±2）℃、转速为150 r/min的条件下，处理pH 7、S^2-和NH⁺₄-N分别为80 mg/L和88 mg/L的水样40 h，硫化物和氨氮的脱除率分别为91.8%和96.6%。     

降氰菌的分离及培养条件初探

从温州某电镀厂的活性污泥中分离纯化、筛选获得降氰菌16株,其中11菌株能耐受2400mg/L的氰化钠.同时,考察了实验条件对11菌株的生长影响,结果表明,11菌株最佳生长pH为8.0,4～18h为对数生长期;果糖对11菌株的生长和降氰有促进作用;50mL含氰液体培养基中,1mol/L的Fe或Cu对11菌株的生长和降氰均有抑制作用;11菌株的最佳降氰液体培养基pH为6.5,4h内的降氰率可达98.57%;8h后CN-残留质量浓度降至501μg/L.     

盐度对活性污泥驯化前后硝化特性的影响

通过批量试验系统研究了盐度对常规活性污泥硝化作用的影响以及污泥在含盐环境中经过驯化后其硝化功能的变化.试验结果表明,硝化菌比亚硝化菌对盐度更敏感,废水中含盐浓度为5 g/L时,对常规活性污泥中的亚硝化菌影响不大,硝化菌会受到一定程度的抑制;当含盐浓度超过10 g/L时,硝化菌和亚硝化菌均会受到严重抑制;含盐浓度大于30 g/L时,亚硝化菌和硝化菌已经完全受到抑制.在污泥驯化初期,耐盐硝化菌群数量较少,比硝化速率较低,硝化产物中亚硝态氮大量积累;随着驯化时间的延长,耐盐硝化菌群数量增加,比硝化速率增加,亚硝态氮累积量减少.污泥经过驯化后,硝化菌群可以逐渐适应高盐环境,在含盐浓度为30 g/L时硝化反应仍能进行.     

纤维素降解细菌筛选及降解特性分析

基于获得高效纤维素降解细菌的目的,通过LB培养基的培养以及刚果红培养基的筛选,从牛粪堆肥中筛选获得2株高效纤维素降解细菌。经鉴定,分别为枯草芽胞杆菌(Bacillus subtilis)和地衣芽胞杆菌(Bacillus licheniformis)。所筛选得到的菌种具有很高的滤纸降解能力,可在6d内使滤纸剧烈崩溃,振摇成均匀糊状;其中,地衣芽胞杆菌的羧甲基纤维素钠酶活峰值在发酵第4天达到峰值(237U/g)。     

Biodegradation of haloacetic acids by bacterial enrichment cultures

Haloacetic acids (HAAs) are toxic organic chemicals that are frequently detected in surface waters and in drinking water distribution systems. The aerobic biodegradation of HAAs was investigated in serum bottles containing a single HAA and inoculated with washed microorganisms obtained from enrichment cultures maintained on either monochloroacetic acid (MCAA) or trichloroacetic acid (TCAA) as the sole carbon and energy source. Biodegradation was observed for each of the HAAs tested at concentrations similar to those found in surface waters and in drinking water distribution systems. The MCAA culture was able to degrade both MCAA and monobromoacetic acid (MBAA) with pseudo-first order rate constants of 1.06 x 10(-2) and 1.13 x 10(-2) l(mg protein)(-1) d(-1), respectively, for concentrations ranging from 10(-5) to 2 mM. The pseudo-first order rate constant for TCAA degradation by the TCAA culture was 6.52 x 10(-3) l(mg protein)(-1) d(-1) for concentrations ranging from 5.33 x 10(-5) to 0.72 mM. The TCAA culture was also able to degrade MCAA with the rate accelerating as incubation time increased. Experiments with radiolabeled HAAs indicated that the 14C was primarily converted to 14CO2 with minor incorporation into cell biomass. The community structure of the enrichment cultures was analyzed by both cultivation-dependent and cultivation-independent approaches. Denaturing gradient gel electrophoresis (DGGE) of the PCR-amplified 16S rRNA gene fragments showed that each of the two enrichment cultures had multiple bacterial populations, none of which corresponded to HAA-degrading bacteria cultivated on HAA-supplemented agar plates. This research indicates that biodegradation is a potential loss mechanism for HAAs in surface waters and in drinking water distribution systems.     

Evaluation of dispersion methods for enumeration of microorganisms from peat and activated carbon biofilters treating volatile organic compounds

To enumerate microorganisms having colonized biofilters treating volatile organic compounds, it is necessary firstly to evaluate dispersion methods. Crushing, shaking and sonication were then tested for the removal of microflora from biofilters packing materials (peat and activated carbon). Continuous or discontinuous procedures, and addition of glass beads had no effect on the number of microorganisms removed from peat particles. The duration of treatment also had no effect for shaking and crushing, but the number of microorganisms after 60 min of treatment with ultrasound was significantly higher than that obtained after 0.5 min. The comparison between these methods showed that crushing was the most efficient for the removal of microorganisms from both peat and activated carbon. The comparison between three chemical dispersion agents showed that 1% Na-pyrophosphate was less efficient, compared with 200 mM phosphate buffer or 1% Na-hexametaphosphate. To optimize the cultivation of microorganisms, three different agar media were compared. Tryptic soy agar tenfold diluted (TSA 1/10) was the most suitable medium for the culture of microflora from a peat biofilter. For the activated carbon biofilter, there was no significant difference between Luria Bertoni, TSA 1/10, and plate count agar. The optimized extraction and enumeration protocols were used to perform a quantitative characterization of microbial populations in an operating laboratory activated carbon biofilter and in two parallel peat biofilters.     

双泥膜法SBR脱氮除磷效果研究

通过实验室小试，以人工模拟生活污水为研究对象，考察了基于反硝化除磷理论开发的双泥膜法SBR工艺的脱氮除磷效果。长期试验结果表明：双泥膜法SBR工艺能使硝化菌和除磷菌各自在最佳的环境中生长，解决了传统工艺中脱氮和除磷的矛盾，节省了碳源和能源，并取得了稳定高效的脱氮除磷效果。在进水C∶N∶P为25∶5∶1，换液比为75%的情况下，系统对COD，TP和NH⁺₄-N的平均去除率分别为89.13%、96.12%和86.78%。     

臭氧-固定化生物活性炭滤池深度处理石化废水的试验研究

针对石化废水中不同特征污染物,采用人工分离筛选去除COD和油工程菌6株、硝化工程菌10株(亚硝化细菌5株、硝化细菌5株)构建高效混合菌群,通过臭氧固定化生物活性炭滤池除污染效能中试研究表明,该系统深度处理石化难降解有机废水是可行的,能同时实现去除COD、油类、NH3 N等污染物的功效,对COD、油类、NH3 N和色度的平均去除率分别为73 0%、90 5%、81 2%和90%,相应的出水分别为33 2mg/L、0 4mg/L、4 5mg/L和10倍,各项指标均达到了国家循环冷却水的用水要求,它的推广应用必将带来显著的环境效益、社会效益和经济效益。     

Effect of Diuron on aquatic bacteria in laboratory-scale wastewater treatment ponds with special reference to Aeromonas species studied by colony hybridization

Six laboratory-scale wastewater treatment ponds were filled with sediment and water obtained from a reference pond (a wastewater treatment plant located in a rural environment at Montel-de-Gelat, Puy-de-D?me, France). They were kept at 20 degrees C, with alternative light and dark periods (12 h-12 h), and fed with raw effluent supplied weekly. Three of them were treated with Diuron (dissolved in DMSO) at a final concentration 10 mg/l, while the other three received only DMSO. Physico-chemical parameters, total bacteria, cultivable bacteria, and Aeromonas spp. were measured periodically until 41 days after the Diuron contamination. Total bacteria were treated with 4,6-diamidino 2-phenylindole (DAPI) and counted by epifluoroscence microscopy. The cultivable bacteria were quantified on plate count agar medium and Aeromonas spp. using colony hybridization. In the contaminated pilots, biochemical oxygen demand (BOD5), chemical oxygen demand (COD), suspended solids (SS), volatile suspended solids (VSS), ammonium, phosphorus, and bacteria increased, but dissolved oxygen decreased. The abundance of total bacteria, cultivable bacteria (multiplied by 30), and Aeromonas spp. increased for two weeks after Diuron introduction, reverting to initial values three weeks later. The percentage of cultivable bacteria relative to total bacteria was 0.2% in controls and 1.2% in treated pilots, while the percentage of Aeromonas spp. relative to cultivable bacteria decreased from 6-10% to 2%. Our results suggest that Diuron, which acts on the photosystem II of phototrophs, supports the development of cultivable bacteria through new carbon sources derived from the decomposition of photosynthetic micro-organisms, but does not specifically support Aeromonas spp.     

Consolidation of degraded ornamental porous limestone stone by calcium carbonate precipitation induced by the microbiota inhabiting the stone

Although it has already been shown that calcareous stone can be consolidated by using a bacterially inoculated culture medium, a more user-friendly method is the in situ application of a sterile culture medium that is able to activate, among the microbial community of the stone, those bacteria with a potential for calcium carbonate precipitation. In order to test this new method for stone consolidation, non-sterilized decayed porous limestone was immersed in sterile nutritional media. Results were compared to those of the runs in which stone sterilized prior to the treatment was used. The effects of the microbial community on stone consolidation were determined by recording the evolution of the culture media chemistry. The treated stone was tested for mechanical resistance and porosity. Results demonstrate that the tested media were able to activate bacteria from the microbial community of the stone. As a consequence of the growth of these bacteria, an alkalinization occurred that resulted in calcium carbonate precipitation. The new precipitate was compatible with the substrate and consolidated the stone without pore plugging. Therefore, a good candidate to in situ consolidate decayed porous limestone is the application of a sterile culture medium with the characteristics specified in the present study.     

斜生栅藻的无菌化培养及其在原油降解中的应用研究

选用组合抗生素结合溶菌酶/SDS预处理方法对斜生栅藻进行无菌化培养,对不同抗生素组合的除菌效果进行了比较。青霉素+庆大霉素对斜生栅藻和附生菌都有强烈的抑制性能,而青霉素+利福平和青霉素+四环素组合除菌不彻底。青霉素+卡那霉素组合能彻底除去附生菌并且对藻的抑制作用最小,可得到无菌斜生栅藻。将斜生栅藻与石油组分降解菌构建菌藻共生体系,未除菌藻的附生菌与石油组分菌之间不能很好地相互适应,使得原油降解效果反而降低。纯栅藻则对石油降解菌表现出非常良好的促进作用,利用除菌后的斜生栅藻与石油组分降解菌构建了一个高效降解原油的菌藻共生体系,充分肯定了微藻无菌化培养的积极意义。