石油污染与食细菌线虫对土壤微生物活性及石油降解的影响

随着经济快速发展,石油及其产品用量增多,石油污染问题日益严重,土壤石油污染治理刻不容缓.研究表明石油污染土壤中存在大量的食细菌线虫,但食细菌线虫在石油污染土壤中的作用还不清楚.本试验采用人工模拟石油污染土壤,通过接种不同密度模式线虫(Caenorhabditis elegans),研究食细菌线虫在石油污染土壤中的功能及其对污染土壤中石油降解及土壤微生物活性的影响.本实验共设6个处理:高温灭菌石油污染土壤(FSP),作为对照处理1;杀灭线虫土壤(S),作为对照处理2;石油污染土壤(SP);石油污染土壤+5条线虫/g干土(SPN5);石油污染土壤+10条线虫/g干土(SPN10);石油污染土壤+20条线虫/g干土(SPN20).研究结果表明:整个试验培养结束时,处理SP、SPN5、SPN10和SPN20的石油残留量比第0天采样时分别降低约60.78%、80.01%、67.63%和66.31%,处理SP、SPN5、SPN10和SPN20的石油残留量比处理FSP分别降低约43.60%、70.68%、52.34%和50.45%,得出接种线虫可以促进污染土壤的石油降解,其中接种5条线虫/g干土的处理促进石油降解效果最好.第7 d采样时,处理SP、SPN5、SPN10、SPN20中脱氢酶酶活性比处理S分别增加约132.76%、115.09%、118.67%和55.81%,表明石油污染可以激活脱氢酶;第14 d时,接种线虫处理SPN5、SPN10和SPN20的脱氢酶活性比未接种线虫处理SP的脱氢酶活性分别增加约5.16%、18.13%和29.56%,表明添加食细菌线虫也促进了土壤相关酶活性.该研究证明食细菌线虫可以在石油污染土壤中刺激微生物的繁殖,增强土壤酶活性,进而促进污染土壤石油的降解.     

堆肥对土壤中石油烃的去除及微生物群落的影响

为研究堆肥对石油污染土壤中不同组分烃的去除作用及土壤微生物群落结构变化的影响,利用重量法和GC-MS测定土壤中总石油烃、烷烃和多环芳烃的含量,采用高通量测序技术研究了堆肥对土壤微生物群落结构和多样性的影响作用.结果表明,向石油污染土壤中施加堆肥进行42d的修复处理,土壤中石油烃、烷烃、多环芳烃去除率分别为（12.4±0.01）%、（10.2±0.01）%、（9.38±0.02）%;自然放置的土壤中3种烃去除率分别为（3.21±0.02）%、（-3.00±0.01）%、（-6.59±0.02）%.自然放置的土壤香农指数、ACE指数和Chao1指数分别为4.30、3489.3和2691.0,加入堆肥进行修复处理后,土壤香农指数、ACE指数和Chao1指数分别增加为5.80、4684.7和3851.8.油污土壤中放线菌门（Actinobacteria）所占丰度由47.3%降低为28.2%,拟杆菌门（Bacteroidetes）丰度由0.78%增加至16.2%.变形菌门（Proteobacteria）丰度为37.4%,修复结束后几乎不变.属水平上,油污土壤中的优势菌属包括原小单孢菌属（promicromonospora）、微小杆菌属（Exiguobacterium）、诺卡氏菌属（Nocardioides）、分支杆菌属（Mycobacterium）、柠檬酸细菌属（Citrobacter）.施入堆肥使土壤中的这些优势菌属丰度降低,土壤中出现氮单胞菌属（Azomonas）、藤黄单胞菌属（Luteimonas）、假鞘氨醇杆菌属（Pseudosphingo bacterium）、紫单胞菌属（Parapedobacter）等新菌属.研究结果表明,与自然放置的土壤相比,向石油污染土壤中施入有机堆肥可有效去除土壤中的石油烃、烷烃和多环芳烃.并使土壤微生物群落结构发生明显变化.     

牵牛花对石油污染盐碱土壤微生物群落与石油烃降解的影响

利用磷脂脂肪酸（PLFAs）生物标记法分析野生型牵牛花（Pharbitis nil （Linn.） Choisy）根际土壤微生物群落结构,探讨牵牛花生长对石油烃污染土壤微生物群落与石油烃降解的影响.结果表明,供试土壤微生物群落中,先后出现了24种PLFAs,包括标记细菌的饱和脂肪酸（SAT）、革兰氏阳性菌（G⁺）的末端支链饱和脂肪酸（TBSAT）、革兰氏阴性菌（G^-）的单不饱和脂肪酸（MONO）和环丙脂肪酸（CYCLO）、真菌的多不饱和脂肪酸（PUFA）和放线菌的中间型支链饱和脂肪酸（MBSAT）等六大类型.PLFAs的主成分分析（PCA）表明,牵牛花根际与对照组（未种牵牛花,CK）土壤微生物群落具有明显的差异,微生物多样性在春季增加83%、夏季增加140%、秋季增加50%;微生物的生物量在春季增加97.6%,夏季增加116.3%,秋季增加60.3%.牵牛花根际与对照组相比土壤中石油烃降解率明显提高,在春、夏、秋季分别提高了7.5%、34.2%和19.7%;并且,在牵牛花生长的不同季节石油烃的降解率有明显的差异,春季为22.3%,夏季为51.8%,秋季为38.0%.相关性分析表明,石油烃降解与土壤微生物总生物量具有中等程度的相关性（|r|=0.75）;与G⁺细菌、甲烷氧化菌的生物量具有高度相关性,相关系数|r|>0.8;与G^-的生物量具有中度相关性,相关系数为|r|=0.74;与真菌的生物量具有低度相关性,相关系数为|r|=0.36,与放线菌没有相关性,相关系数为|r|<0.30.     

二维电场中微生物群落动态及混合有机物降解特征

以正十六烷、环十二烷和芘组成的混合有机污染物为研究对象,在二维对称电场修复平台上,采用电动-微生物联合修复(BIO-EK)的方式,通过分析微生物群落时空动态及混合有机物降解的时空特征,探讨了二维电场中电场强度、微生物群落和污染物降解特征之间的相关性.结果表明,BIO-EK处理过程中,土壤pH和温度在时间和空间上均没有发生明显的变化,微生物群落结构和数量均主要随处理时间而变化,未发生明显的空间上的变化;电场的施加可提高微生物数量,对污染物降解有积极影响,相对于单独的微生物(BIO)和单独的电动(EK)处理,BIO-EK对污染物处理效率最高;不同类型污染物去除率与电场强度均呈正相关.     

Dynamic changes in microbial activity and community structure during biodegradation of petroleum compounds： A laboratory experiment

With 110-d incubation experiment in laboratory,the responses of microbial quantity,soil enzymatic activity,and bacterial community structure to different amounts of diesel fuel amendments were studied to reveal whether certain biological and biochemical characteristics could serve as reliable indicators of petroleum hydrocarbon contamination in meadow-brown soil,and use these indicators to evaluate the actual ecological impacts of 50-year petroleum-refining wastewater irrigation on soil function in Sbenfu irrigation area.Results showed that amendments of≤1000 mg/kg diesel fuel stimulated the growth of aerobic beterotrophic bacteria,and increased the activity of soil dehydrogenase,hydrogenperoxidase,polypbenol oxidase and substrate-induced respiration.Soil bacterial diversity decreased slightly during the first 15 d of incubation and recovered to the control level on day 30.The significant decrease of the colony forming units of soil actinomyces and filamentous fungi can be taken as the sensitive biological indicators of petroleum contamination when soil was amended with≥5000 mg/kg diesel fuel.The sharp decrease in urease activity was recommended as the most sensitive biochemical indicator of heavy diesel fuel contamination.The shifts in community structure to a community documented by Sphingomonadaceae withinα-subgroup of Proteobacteria could be served as a sensitive and precise indicator of diesel fuel contamination.Based on the results described in this paper,the soil function in Shenfu irrigation area was disturbed to some extent.     

土壤微生物电化学系统降解四环素的机理

以四环素为研究对象,构建土壤微生物电化学系统(SMES),避光恒温培养58d后进行采样分析,研究了四环素降解、土壤理化性质、酶活性和三域微生物之间的内在联系,以揭示四环素在SMES中的生物降解机理.结果表明,采用SMES处理后,四环素降解率从52％显著提升至70％.与对照处理相比,脱氢酶活性在SMES中显著提升了144...     

营养盐对湄洲湾海洋细菌生长及降解石油烃的影响

测定从湄洲湾海域分离的烃细菌在添加和不添加N、P营养盐的海水培养基的生物量及对原油和纯烃的降解作用.结果表明,营养盐对不同菌株的生长及代谢有不同的影响.添加N、P营养盐,试验菌PF-6（Pseudomonas fluorescens 6）的生物量增大,而PA-32（P.aeruginosa 32） 的生物量却减少.在初始原油浓度均为1g/L的摇瓶试验中,添加N、P营养盐培养6d,PF-6菌与PA-32菌的除油率分别为25%及26%,而不添加N、P营养盐时,PF-6菌与PA-32菌的除油率分别为21.4%及36.3%.经气相色谱测定,在以正十六烷和萘两种纯烃组成的培养基,营养盐对两个菌株降解正十六烷的影响仍然不同,但在不添加N、P营养盐时,两个菌株对萘均有较高的降解率.无需添加N、P营养盐能正常生长并降解石油烃的菌株,在海洋油污的生物修复中具有应用前景.     

Effects of butachlor on microbial enzyme activities in paddy soil

This paper reports the influences of the herbicide butachlor(n-butoxymethl-chloro-2‘,6‘-diethylacetnilide)on microbial respiration,nitrogen fixation and nitrification,and on the activities of dehydrogenase and hydrogen peroxidase in paddy soil.The results showed that after application of butachlor with concentrations of 5.5μg/g dried soil,11.0μg/g dried soil and 22.0μg/g dried soil,the application of butachlor enhanced the activity of dehydrogenase at increasing concentrations.The soil dehydrogenase showed the highest activity on the 16th day after application of 22.0μg/g dried soil of butachlor.The hydrogen peroxidase could be stimulated by cutachlor.The soil respiration was depressed within a period from several days to more than 20 days,depending on concentrations of butachlor applied.Both the nitrogen fixation and nitrification were stimulated in the beginning but reduced greatly afterwards in paddy soil.     

典型油田区油污土壤微生物群落区域性分布研究

石油污染改变土壤微生态环境,驱动了土壤微生物群落结构的演替与进化.为了深入探究油田区油污土壤中微生物群落分布特征,揭示区域性的土壤微生物群落结构成因,采用Miseq平台的16S rDNA扩增子测序技术分析了辽河油田和大庆油田区6组共计18个土壤样品的微生物群落多样性及结构组成,并结合土壤环境因子指标剖析了群落结构成因,进而预测了具有石油代谢能力的功能菌属.结果表明,石油含量随着距井口距离增加而减少,石油的空间分布特征是影响微生物群落结构变化的关键因子,6组土壤样品的OTU分属于49门、131纲、169目、328科和564属,微生物种群多样性随着污油浓度的增加而减小;两油田共有5个相同优势菌门,2种优势菌属;辽河油田区独特优势菌门为Saccharibacteria门,优势菌属为微枝形杆菌属（Microvirga）、分支杆菌属（Mycobacterium）和Defluviicoccus属;大庆油田独特优势菌门为拟杆菌门（Bacteroidetes）,独特优势菌属包括盐单胞菌属（Halomonas）、食烷菌属（Alcanivorax）和海杆菌属（Marinobacter）等.冗余分析（RDA）结果表明污油组成是微生物群落差异性分布的决定性因素,胶质的高含量与强毒性诱导辽河油田区微生物群落获得较强的胁迫抗性;同时区域生态环境背景差异也是影响微生物群落整体胁迫抗性的重要因子.结合PICRUSt分析预测,共发现2种辽河油田区和5种大庆油田区石油功能降解优势菌属,为石油降解功能菌剂的种质资源的高效开发提供目标菌株.     

除草剂硝磺草酮对土壤微生物生态效应研究

采用室内培养试验,研究了硝磺草酮对土壤酶活性和微生物群落功能多样性的影响.结果表明,土壤过氧化氢酶和蔗糖酶活性随硝磺草酮浓度增加呈现先增加后降低的趋势,但比对照处理都有所升高,分别增加了22.6%~41.0%和3.4%~54.2%,且在硝磺草酮浓度为50mg/kg时达到最大.与之相反,尿酶活性则降低了12.0%~18.6%,但差异性不明显(P > 0.05).施用一定浓度的硝磺草酮激活了土壤微生物活性,微生物群落丰富度、均匀性和多样性都呈增长趋势,单孔的平均颜色变化率(AWCD)值和利用速率均随硝磺草酮浓度增加而增大,微生物群落对碳水化合物类、氨基酸类、多聚物类、羧酸类、胺类和酚酸类利用率整体上均有所提高,与对照相比,最大增幅分别达到5.3、1.0、4.4、3.2、0.2和6.8倍,但不同浓度硝磺草酮处理下土壤微生物在利用碳源的类型上是存在一定的差异.     

微生物胞外电子传递过程强化机制及污染物高效转化

微生物水处理技术因运行成本低、处理量大、环境影响小等优点,被广泛的用于市政污水和工业废水的处理.微生物水处理的本质过程是生物催化氧化,涉及不同微生物种群间物质、能量和电子传输过程,而微生物胞外电子传递过程是影响其处理效率的关键因素之一.本研究立足于微生物氧化的原理,从介体材料强化、光电磁强化及微生物电化学强化等角度,系统论述了铁基材料、碳基材料、光、电、磁等对微生物电子传递过程的影响与机制.在此基础上,本论文总结了微生物电化学系统原理,分析了该系统中各类强化材料、强化技术对微生物胞外电子传递的影响,揭示了强化条件下污染物高效转化的作用机制,介绍了基于各种强化原理的系统构建因素及应用现状,并展望了该技术的发展趋势及存在的挑战.     

微生物菌剂对砒砂岩土壤的改良作用

为了研究微生物菌剂改良砒砂岩土壤特性的效果,以植物促生细菌Bacillus halotolerans P75、Sinorhizobium meliloti D10、Bacillus megaterium H3和Bacillus subtilis HB01为原料制备4种液体单一菌剂和1种复合菌剂CB,分别按5%（体积质量比）接种到砒砂岩土壤中,种植紫花苜蓿和柠条,进行盆栽试验.通过分析植物生物量、土壤pH值、有机质、氮磷钾含量、酶活性和水稳性团聚体等指标来评价微生物菌剂对砒砂岩土壤的改良效果.结果表明,与不接菌的对照组相比,接菌处理能使紫花苜蓿显著增产18.6%~45.6%、柠条显著增产24.1%~46.7%,降低土壤pH值,促使土壤中的有机质、铵态氮、硝酸盐氮、速效磷、速效钾含量不同程度的增加,提高了土壤团聚体的水稳性,且接种复合菌对砒砂岩土壤的改善效果要优于单一菌株的效果.接种供试菌株能够改善砒砂岩的土壤质量,同时促进植物生长,其中复合菌对砒砂岩土壤的改良效果更为突出.     

Effect of rhizosphere on soil microbial community and in-situ pyrene biodegradation

To access the influence of a vegetation on soil microorganisms toward organic pollutant biogegration, this study examined the rhizospheric effects of four plant species (sudan grass, white clover, alfalfa, and fescue) on the soil microbial community and in-situ pyrene (PYR) biodegradation. The results indicated that the spiked PYR levels in soils decreased substantially compared to the control soil without planting. With equal planted densities, the efficiencies of PYR degradation in rhizosphere with sudan grass, white clover, alfalfa and fescue were 34.0%, 28.4%, 27.7%, and 9.9%, respectively. However, on the basis of equal root biomass the efficiencies were in order of white clover >> alfalfa > sudan > fescue. The increased PYR biodegradation was attributed to the enhanced bacterial population and activity induced by plant roots in the rhizosphere. Soil microbial species and biomasses were elucidated in terms of microbial phospholipid ester-linked fatty acid (PLFA) biomarkers. The principal component analysis (PCA) revealed significant changes in PLFA pattern in planted and non-planted soils spiked with PYR. Total PLFAs in planted soils were all higher than those in non-planted soils. PLFA assemblages indicated that bacteria were the primary PYR degrading microorganisms, and that Gram-positive bacteria exhibited higher tolerance to PYR than Gram-negative bacteria did.     

黄土地区石油类污染物的径流污染模拟及模型预测

在土壤石油污染负荷为721－16060mg/kg、暴雨强度为 0.5－1.70mm/min的条件下进行了室内径流污染模拟试验研究．结果表明,随土壤石油污染负荷的提高,产流量与释放至水中的污染物浓度均增大,但产沙量降低;随暴雨强度的增大,产流、产沙、产污的强度均提高．试验条件下释放于水中的石油污染物浓度可高达1.56－15.6mg/L,由此进一步表明,暴雨径流对水体造成的石油污染不可轻视．在试验研究基础上,建立了径流污染过程的稳态产流、产沙、产污模型;应用该组模型对土壤石油污染负荷为 7050mg/kg和不同雨强下的径流污染进行了预测,预测结果与试验模拟结果吻合良好     

Effect of acetate on lead toxicity to microbial biomass in a red soil

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生物炭输入土壤对其石油烃微生物降解力的影响

以木屑和麦秆为原料,在300℃和500℃下热解制备生物炭,分析了生物炭输入对大港油田污染土壤中总石油烃及其组分(正链烷烃n C8~C40和16种PAHs)微生物降解效果的影响.结果表明,生物炭输入强化了土壤中总石油烃及其组分的生物降解.生物炭原料的选取对烷烃降解影响显著,对PAHs影响较小;高温制备生物炭对污染物降解的强化效果较好,这归因于生物炭表面性质和降解微生物种类的不同.土壤中加入生物炭后,低环PAHs的降解效率显著高于高环PAHs.添加典型的土壤易分解有机质(葡萄糖)产生正激发作用,导致生物炭矿化,促进了烷烃降解,抑制了PAHs的去除.可见,生物炭输入可有效促进石油烃的微生物降解,对修复石油污染土壤起正效应.     

石油烃对虾夷马粪海胆胚胎发育及四种酶活力的影响

通过虾夷马粪海胆胚胎暴露实验,研究了常见石油烃零号柴油对海胆胚胎发育时间以及发育时期内海胆胚胎的SOD、CAT、GPx及谷胱甘肽转移酶(GST)四种酶活性的影响。结果表明,不同浓度的零号柴油水溶组分对海胆胚胎发育的各个时期具有不同程度的延迟效应,低浓度对海胆胚胎中的四种酶表现出不同程度的诱导效应;四种酶活性的变化程度比较,对GPx和GST酶活力的影响要明显大于对CAT和SOD的影响;随着浓度的不断增大,四种酶活性受到不同程度的抑制作用,高浓度对酶活性的抑制作用尤为显著;囊胚期对影响最为敏感,可作为生物标志物。     

Eco-toxicity of petroleum hydrocarbon contaminated soil

Total petroleum hydrocarbons (TPH) contaminated soil samples were collected from Shengli Oilfield of China. Toxicity analysis was carried out based on earthworm acute toxicity, plant growth experiment and luminescent bacteria test. The soil was contaminated bypetroleum hydrogcarbons with TPH concentration of 10.57%. With lethal and sub-lethal rate as endpoint, earthworm test showed that the LD50 (lethal dose 50%) values in 4 and 7 days were 1.45% and 1.37% respectively, and the inhibition rate of earthworm body weight increased with higher oil concentration. TPH pollution in the soil inhibited seed germination in both wheat and maize experiment when the concentration of petroleum was higher than 0.1%. The EC50 (e ective concentration 50%) for germination is 3.04% and 2.86% in maize and wheat, respectively. While lower value of EC50 for root elongation was to be 1.11% and 1.64% in maize and wheat, respectively, suggesting higher sensitivity of root elongation on petroleum contamination in the soil. The EC50 value in luminescent bacteria test was 0.47% for petroleum in the contaminated soil. From the experiment result, it was concluded that TPH content of 1.5% is considered to be a critical value for plant growth and living of earthworm and 0.5% will a ect the activity of luminescent bacteria.     

石油污染物在土壤中运移的数值模拟初探

针对大庆石油管理局宋芳屯油田开发建设的实际情况，分析了石油污染物在土壤中运移的规律，并建立了非饱和带污染物迁移和地下水运动模型。通过求有限元数值解，利用该模型对石油污染物在土壤中的迁移情况进行了模拟预测，结果显示了石油污染物可能对土壤造成污染的范围和影响程度。     

Cometabolic microbial degradation of trichloroethylene in the presence of toluene

Trichloroethylene (TCE), a common groundwater pollutant, was cometabolized by microorganisms in the presence of toluene as a growth substrate. The effect of concentrations of toluene and TCE and temperature on biodegradation was discussed. Acclimated microorganisms degraded TCE after a lag period of 5 to 22 h depending on toluene concentrations. Approximately 60%, 90% and 04% of TCE were degraded at toluene to TCE concentration ratios of 23 : l, 115 : 1 and 230: l, respectively. At a TCE concentration of 1.46 μg/ml, 80% of TCE and 98.4% of toluene were removed. But less degradation of TCE and toluene was observed when TCE concentration was above 48.8 μg/ml. The lag time of TCE decreased and the TCE biodegradation rates increased with the increase of temperature.