低营养水体中芽孢杆菌降解有机氮的研究

芽孢杆菌具有降解有机氮的功能,但在养殖水体等低营养水体中,其降解效果可能受到影响.为研究低营养水体中有机氮的降解情况,通过模拟凡纳滨对虾中间培育过程配制低营养水体,分别接种芽孢杆菌NT9和YB3(NT9水体和YB3水体),然后研究水体中微生物的生长与有机氮的降解情况,并构建数学模型进行分析.结果显示,起始接种量为10×10~5 cfu·mL~(-1)时,NT9水体总菌量呈下降趋势,平均为(3.46×10~5±2.39×10~5) cfu·mL~(-1),YB3水体总菌量则上升到(25.43×10~5±8.84×10~5) cfu·mL~(-1),但均高于未接种的对照水体.NT9水体和YB3水体的有机氮降解率显著高于对照水体(p0.05),分别提高50.28%和119.41%,降解速率也分别提高65.22%和121.74%.对照水体、NT9水体和YB3水体单位菌量的有机氮降解效率分别为1.238、1.649和1.904 mg·L~(-1),降解模型分别为y=-6.40+1.39x_1+1.45x_2、y=2.11+8.21x_3-0.64x_4-1.26x_1x_3-0.32x_2x_4和y=1.73+6.11x_2(x_1、x_2、x_3、x_4分别表示总菌量、总菌增量、有机氮含量和时间).研究表明,在低营养水体中接种芽孢杆菌有利于有机氮的降解,但不同的菌株具有不同的降解模式,菌株YB3为能够适应低营养水平、增殖能力较强的菌株,可以更有效地促进有机氮的降解,提高降解效率.     

耐冷嗜碱蒙氏假单胞菌H97的鉴定及其好氧反硝化特性

结合形态、磷脂脂肪酸及16S rRNA基因序列分析鉴定了分离自贵州冬水田的好氧反硝化菌株H97,采用模拟废水探讨了不同温度、接菌量、C/N、初始p H和碳源种类对菌株H97反硝化能力的影响.结果表明,菌株H97为蒙氏假单胞菌(Pseudomonas monteilii),目前国内外尚无蒙氏假单胞菌具有耐冷嗜碱好氧反硝化作用的研究报道,是对好氧反硝化菌的补充;该菌的最适脱氮条件为15℃、p H 9.0、C/N 15、接种量1.5×106CFU·(100 m L)-1,碳源丁二酸钠.此外,菌株H97在p H为7.0~11.0时,均表现出良好的脱氮能力,对硝酸盐氮和总氮的去除率分别可达91.21%和79.10%以上,初始p H为12.0时,对硝酸盐氮和总氮的去除率仍分别达64.75%和36.78%,表现出了较强耐碱能力.在初始硝酸盐氮浓度为50.0 mg·L-1和最适脱氮条件下,菌株H97在48 h内对硝酸盐氮和总氮的去除率可达97.69%和96.32%.同时,H97对温度适应范围广,在15~40℃均具有较强好氧反硝化能力,是1株耐冷嗜碱好氧反硝化细菌,在碱性氮污染水体处理中具有较好的应用潜力.     

一株异养硝化细菌的分离鉴定及其脱氮效果

该研究从富营养化景观水体中分离出40株细菌,利用PCR-RFLP(限制性片段长度多态性)技术确定了11种不同的基因型,对11种不同基因型菌株氨氮(NH4+-N)和总氮(TN)的去除效果进行研究,发现其中1株菌在24 h内NH4+-N的去除率达87.7%,TN去除率达81.7%,而且该菌从培养体系中完全去除的氮明显多于同化到细胞内的氮。进一步对该菌进行分子鉴定,确定了其种属特征,并命名为Klebsiella sp.HLNR02。利用菌株HLNR02处理模拟富营养化废水,结果发现其在48 h内对模拟废水中的NH4+-N和TN去除率达85.3%和77.3%,C/N比会影响菌株的解氮效果。     

耐冷亚硝酸盐型反硝化细菌Pseudomonas putida Y-12脱氮特性

研究了温度、转速、pH、接种量、碳源和亚硝酸盐氮浓度对耐冷反硝化细菌Pseudomonas putida Y-12去除亚硝酸盐氮和总氮的影响,在此基础上,初步考察了反硝化脱氮产物中温室气体(CO2和N2O)的含量,探索了对富营养化景观水体中亚硝酸盐氮和总氮的去除能力.结果表明:Y-12菌株去除亚硝酸盐氮和总氮的最佳碳源为柠檬酸钠,在15℃和pH为7.2条件下,该菌以柠檬酸钠为唯一碳源对亚硝酸盐氮和总氮去除率分别达到了100%和81.8%;反硝化脱氮产生了19559.82 mg·m-3CO2和0.11 mg·m-3N2O;在15℃,150 r·min-1摇床培养条件下,48 h内菌株Y-12对景观水体中亚硝酸盐氮和总氮去除率分别达到了37.7%和51.5%,表明该菌在初春及晚秋水体的低温时段具有较好的应用潜力.     

1株海洋异养硝化-好氧反硝化菌的分离鉴定及其脱氮特性

以海水为基质,采用传统的微生物分离纯化方法,从海底沉积物中分离筛选得到1株耐盐异养硝化-好氧反硝化细菌y5,经形态、生理生化特性以及16S rRNA基因序列分析,鉴定该菌为克雷伯氏菌(Klebsiella sp.).对其脱氮特性及影响因素进行了研究,结果表明,菌株y5的最佳碳源为柠檬酸三钠,最适p H值为7.0,最适C/N为17.菌株均能以NH4Cl、Na NO2和KNO3为唯一氮源进行反应,36 h的去除率分别为77.07%、64.14%和100%.3种氮源共存时,36 h的去除率达到100%.表明菌株y5在高盐废水中具有独立高效的异养硝化和好氧反硝化作用.     

一株好氧反硝化菌的分离鉴定及脱氮条件研究

好氧反硝化菌在生物脱氮中有很好的应用前景,文章旨在从环境中分离具有高效脱氮能力的反硝化菌,对其进行分类及反硝化特性研究。以对硝酸盐氮的去除能力为主要指标,从对虾养殖池底泥分离筛选出有效去除硝酸盐氮的好氧反硝化菌,通过生理生化特性及16S r DNA序列分析对菌株进行初步鉴定。在好氧条件下,研究不同碳源、氮源、温度、p H值对该菌总氮去除率的影响。试验结果表明菌株S4在以柠檬酸钠为唯一碳源,30℃,100 r/min振荡培养24 h和36 h,硝酸盐氮去除率分别达到60%和72%。其16S r DNA序列与海源菌的同源性达98%,结合生理生化特点,初步确定菌株S4属于海源菌属(Idiomarina sp)。最适碳源、氮源、分别为乙酸钠和牛肉膏,最适培养温度为20℃,最适p H值为8,在最适条件下总氮去除率达80%。     

反硝化细菌的筛选及菌藻共培养体系除氮特性

为提高养殖废水的除氮效果,该文从精养池塘水体中分离出1株高效好氧反硝化细菌TC-1,通过形态学观察、生理生化试验并结合16S r DNA序列分析对反硝化细菌TC-1进行了鉴定。选取椭圆小球藻与反硝化细菌TC-1为研究对象,运用菌藻共培养法研究了碳源种类、总接种量、初始p H值、培养温度、摇床转速对菌藻共培养体系除氮性能的影响,并应用Box-Behnken中心组合进行3因素3水平的实验设计对培养条件进行优化。结果表明,菌株TC-1被鉴定为Pseudomonas toyotomiensis TC-1。菌藻共培养体系适宜的脱氮条件为:在以丁二酸钠为碳源,总接种量2%,初始pH 7.04,培养温度30℃,摇床转速169 r/min的条件下,亚硝氮和总氮去除率分别达到99.14%和97.06%,优于椭圆小球藻单藻和反硝化细菌TC-1单菌的的除氮效果,显示了其在废水脱氮中具有良好的应用前景。     

废水脱氮与沼气脱硫耦联菌株的驯化和分离

在不接种污泥、接种厌氧污泥和接种好氧污泥的条件下,采用鼓泡反应器研究猪场废水脱氮与沼气脱硫耦联反应器的启动及关键微生物.试验前期(第26 d前),接种污泥反应器的脱氮脱硫率为50%～64%,而不接种污泥反应器的脱氮脱硫率只有11%～14%.到驯化结束时(第56 d),3个反应器的脱氮效率为90%左右,脱硫效率达到70%以上.结果表明,不接种污泥反应器经过一段时间驯化后也可以达到同样的脱氮脱硫效果,只是启动时间比接种污泥的反应器稍长.在反应器启动期间,于不同时段分别进行了微生物种群动态变化检测,结果显示微生物种群数量变化与3个反应器的脱氮脱硫效果变化趋势基本一致.在驯化成功的反应器中,分离筛选出氮硫去除率同时达到60%以上的菌株2株,初步鉴定为脱氮硫杆菌(Thiobacillus denitrificans)和假单胞菌属(Pseudomonas)细菌.     

耐盐异养硝化菌qy18和中度嗜盐异养硝化菌gs2的脱氮特性与耐盐性研究

研究了两株异养硝化菌qy18和gs2的脱氮特性和耐盐范围。筛选出一株耐盐异养硝化菌qy18和一株中度嗜盐异养硝化菌gs2,通过形态观察、生理生化试验和16S rRNA序列分析,分别确定其为假单胞菌属(Pseudom onas)和盐单胞菌属(Halom onas)。在48 h内菌株qy18和菌株gs2的NH4-N去除率分别为98.51%和96.10%,TN去除率分别为83.57%和81.46%,COD去除率分别为54.13%和45.0%,两株菌可以同时利用有机碳源和NH4-N为底物进行硝化反应和同步脱氮。两株菌的最大NH4-N去除速率分别为5.79 mg/(L.h)和5.50 mg/(L.h),高于目前已报道的异养硝化菌的NH4-N去除速率。培养期间没有检测到NO3-N,NO2-N最大积累量分别为0.02 mg/L和3.79 mg/L,推测菌株qy18具有特殊的NH4-N代谢模式。两株菌适宜生长的盐度范围分别为0%~4%和2%~10%。同时发现了在盐度不断升高的过程中,NH4-N去除所受盐度的抑制要早于菌体生长所受盐度的抑制。     

原污泥直接启动厌氧氨氧化装置试验研究

建立2组火山岩填料生物滤池,一组接种污水厂原污泥,另一组接种厌氧氨氧化菌,以人工配水为进水,控制反应器内温度在30~34℃,pH在7.5~8.0,经过前期的调整和后期的运行,达到良好的脱氮效果。接种厌氧氨氧化菌反应器氨氮去除率和亚硝态氮去除率可达90%;接种原污泥反应器氨氮和亚硝态氮去除率可达95%。接种原污泥反应器具有较强的耐冲击负荷能力,可以更好的持留厌氧氨氧化菌。     

EDTA-enhanced phytoremediation of lead contaminated soil by Bidens maximowicziana

Phytoremediation is a potential cleanup technology for the removal of heavy metals from contaminated soils.Bidens maximowicziana is a new Pb hyperaccumulator,which not only has remarkable tolerance to Pb but also extraordinary accumulation capacity for Pb.The maximum Pb concentration was 1509.3 mg/kg in roots and 2164.7 mg/kg in overground tissues.The Pb distribution order in the B. maximowicziana was:leaf>stem>root.The effect of amendments on phytoremediation was also studied.The mobility of soil Pb and the Pb concentrations in plants were both increased by EDTA application.Compared with CK(control check),EDTA application promoted translocation of Pb to overground parts of the plant.The Pb concentrations in overground parts of plants was increased from 24.23-680.56 mg/kg to 29.07-1905.57 mg/kg.This research demonstrated that B.maximowicziana appeared to be suitable for phytoremediation of Pb contaminated soil,especially,combination with EDTA.     

Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day, Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury （HgT）, combinative mercury （Hgc） and gaseous mercury （HgG）, and content of total organic carbon （TOC）, but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average Hgv of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with Hgc in profile 1992 and 2000, and Hgv was significantly correlated with Hg6 in profile 1996. HgG/Hgv ratio in profile samples decreased in the order of （HgG,/HgT）1992〉（HgG/HgT）1996〉〉（HgG/HgT）2000. A simple outline of Hg release in landfill could be drawn： with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf〉root〉stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants （e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum） are capable of enriching more Hg than herbaceous plants.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Enzymatic oxidation of aqueous pentachlorophenol

The influence of the nonionic surfactant Tween 80 on pentachlorophenol （PCP） oxidation catalyzed by horseradish peroxidase was studied. The surfactant was tested at concentrations below and above its critical micelle concentration （CMC）. Enhancement of PCP removal was observed at sub-CMCs. The presence of Tween 80 in the reaction mixture reduced enzyme inactivation which occurred through a combination of free radical attack and sorption by precipitated products. A simple first-order model was able to simulate time profiles for enzyme inactivation in the presence or absence of Tween 80. At supra-CMCs, the surfactant caused noticeable reductions in PCP removal, presumably through micelle partitioning of PCP which precluded the hydrophobic PCP molecule from interacting with the enzyme.     

Decomposition of alachlor by ozonation and its mechanism

Decomposition and corresponding mechanism of alachlor, an endocrine disruptor in water by ozonation were investigated. Results showed that alachlor could not be completely mineralized by ozone alone. Many intermediates and final products were formed during the process, including aromatic compounds, aliphatic carboxylic acids, and inorganic ions. In evoluting these products, some of them with weak polarity were qualitatively identified by GC-MS. The information of inorganic ions suggested that the dechlorination was the first and the fastest step in the ozonation of alachlor.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

基于综合水质标识指数和对应分析法的校园河道水质评价

以三峡大学的校园河道求索溪为研究对象,利用综合水质标识指数法确定求索溪水质类别,分析其水质时空变化规律,并利用对应分析法得出求索溪中不同监测点的主要污染因子.研究结果表明:求索溪整体的综合水质标识指数为7.423,整体水质为劣V类(地表水环境质量标准GB 3838-2002)且黑臭.从时间变化来看,求索溪4月份的水质最差,5月份次之,4、5月份所有监测点的水质都劣于V类且黑臭;8月份水质最好,水质为Ⅳ类;从空间分布来看,8个监测点综合水质标识指数均超过6.0,水质为劣V类,其中6号监测点的水质相对最好,监测点3号的水质相对最差;对应分析法得出求索溪的整体水体污染程度受总氮因子的影响最大,其次为总磷.该研究拟为求索溪及类似校园河道的水环境治理研究提供基础依据和参考.     

Effects of chitosan on growth of an aquatic plant (Hydrilla verticillata) in polluted waters with different chemical oxygen demands

Effects of chitosan on a submersed plant, Hydrilla verticillata, were investigated. Results indicated that H. venicillata could prevent ultrastructure phytotoxicities and oxidativereaction from polluted water with high chemical oxygen demand (COD). Superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents in H. verticillata treated with 0.1% chitosan in wastewater increased with high COD (980 mg/L) and decreased with low COD (63 mg/L), respectively. Ultrastructural analysis showed that the stroma and grana of chloroplast basically remained normal. However, plant cells from the control experiment (untreated with chitosan) were vacuolated and the cell interval increased. The relict of protoplast moved to the center, with cells tending to disjoint. Our findings indicate that wastewater with high COD concentration can cause a substantial damage to submersed plant, nevertheless, chitosan probably could alleviate the membrane lipid peroxidization and ultrastructure phytotoxicities, and protect plant cells from stress of high COD concentration polluted water.