生物强化修复石油污染土壤

筛选高效石油降解菌,考察菌株的降解性能及降解机理,进行花盆模拟高效外源菌强化修复石油污染土壤实验,在降解后期添加激活剂H2O2以及木屑来试图改善微生物的修复环境,减缓微生物的衰亡,并考察修复效果。结果表明,菌株L-1的降解效果较好,其对pH和温度有较大范围的适应性,能分泌较多的表面活性物质,细胞疏水性较强。将其应用于土壤修复中,经过50 d的修复,石油残留率达到50.6%左右,生物强化比自然修复残留率降低了8%左右。在第45天添加激活剂能有效改善修复效果,70 d时添加外源菌的土样最小石油残留率达到37.9%。     

降解菌ZQ5与紫茉莉对芘污染土壤的联合修复

在温室盆栽条件下,通过单独种植紫茉莉、单独接种多环芳烃(PAHs)模式化合物芘的专性降解菌ZQ5和两者的联合修复的3种处理,对芘污染土壤的修复效果进行了研究。结果表明,经90 d修复后,植物-微生物联合修复可将人工污染土壤中的芘降解81.1%,将石油污染土壤中的芘降解50.3%,其修复效率明显高于其他2种处理,是紫茉莉修复的1.98倍,是降解菌ZQ5修复的1.39倍。ZQ5的不同接菌量对于修复60 d后的降解率影响不大。外源生物修复条件下,10～20 cm土壤的修复效率要高于5 cm土壤;自然降解条件下,5 cm土层降解率略高于其他土层。     

柴油降解菌的筛选、菌群构建及其对柴油和十五烷的降解机理

针对柴油污染土壤生物修复技术效率低的问题,通过构建高效降解菌群修复柴油污染的土壤,采用组合优化和正交实验构建最佳组合与接种比例的菌群,并研究其柴油降解特性。结果表明,通过筛选、鉴定并命名的4株柴油降解菌为Bacillus sp. VOC18-L1、 Enterococcus faecalis-L2、 Lysinibacillus-L3、 Rhodococcus equi-L4;当4株菌接种比例为3∶1∶3∶4,pH=7.0,30℃,转速150 r·min~(-1)时,柴油降解的效果最佳,14 d对7.0 mL·L~(-1)的柴油降解率达到89.0%。通过气相色谱质谱联用仪(GC-MS)检测柴油降解产物,发现该混合菌株能将柴油中的烷烃降解为短链烷烃,最终转化为小分子物质。同时利用KEGG数据库获得代谢丰度图并初步预测每种菌的功能,根据微生物多样性测试结果,进一步证明了混合菌对柴油完全降解的效果优于单种菌种。通过人工构建的微生物菌群可以有效地应用于柴油污染土壤的修复。     

固定化微生物修复石油污染土壤影响因素研究

针对石油污染土壤修复,利用实验室已筛选的高效石油降解单菌SM-3,以天然有机材料为载体,吸附法制备固定化微生物。将游离与固定化微生物应用于室内花盆模拟修复石油污染土壤,对C/N/P、微生物投加量、石油含量、氧化剂和表面活性剂设计5因素4水平正交实验,探讨不同修复时期各影响因素的重要性顺序,最佳条件下各菌株的修复效果。结果表明,不同微生物在不同降解时期,各影响因素的重要性会发生变化;经过21 d的修复,固定化单菌SM-3石油降解率为22.77%,修复过程中,接种量是最重要的影响因素,营养元素N、P投加影响较大,表面活性剂和氧化剂影响次之。     

石油降解菌的筛选优化及其对油污土壤的修复特性

分别以牛肉膏蛋白胨-布氏哈斯培养基、蓝色凝胶培养基作为初筛和复筛培养基,从石油污染土壤中筛选出2株可产生微生物表面活性剂的石油烃降解菌。并将菌株投加到油污土壤中进行修复研究,考查了不同影响因素对修复效果的影响。研究结果表明,(1)2株菌对中度石油污染土壤有较好的修复效果,向油污土壤中直接投加菌株修复70 d时对石油烃的去除率为52%;(2)向油污土壤中投加降解菌并同时补充氮营养液,修复70 d时对土壤中总石油烃的去除率可达到75%;对土壤中正构烷烃的去除率为66%;(3)与土壤的含水率及土著菌的降解效果相比,向油污土壤中投加降解菌以及补充氮磷营养液是影响石油污染土壤修复效果的关键因素。     

利用不同组分原油逐级驯化筛选高效石油烃降解混菌

利用不同组分原油逐级驯化的方法对克拉玛依油田的石油污染土样进行石油烃降解混菌的富集驯化,得到一组对稀油和稠油均具有高效降解能力的混菌M3。与采用单一原油驯化方法相比,混菌M3对稀油和稠油的降解率分别提高了12.5%和22%。该混菌具有较强的产表面活性剂的能力,能够使发酵液的表面张力从69.8 mN·m~(-1)降至27.9 mN·m~(-1)。通过混菌M3的生长条件优化实验得出:在温度30℃、pH 7～8、盐度1%、氮源选择尿素的条件下,混菌M3对原油的降解率最高。通过考察混菌M3在污染土壤中对原油的降解效果,发现:在修复期间,土壤脱氢酶呈先升高后降低的趋势;混菌M3可使饱和烃组分增加,并使芳香分、胶质和沥青质组分降低,对重质组分具有较好的降解效果。混菌M3的加入改变了原油性质,促进了土壤中原油的降解,经过56 d修复,土壤中原油降解率达到55.3%。     

植物-微生物联合修复石油污染土壤的实验研究

筛选高效石油降解菌并考察菌株的石油降解能力,通过植物-微生物联合修复石油污染土壤室内实验,在修复过程中测定了土壤中细菌和固氮菌,碱解氮、速效磷和速效钾的含量变化,同时采用傅立叶变换离子回旋共振质谱(ESI FT-ICR MS)考察了植物-微生物联合修复效果。结果表明,菌株3#、4#的生长适应性较强,其混合菌的降解效果最好,将其混合菌液与植物进行植物-微生物联合修复不同浓度的石油污染土壤,经过150 d的温室降解,最高降解率达到73.47%。ESI FT-ICR MS分析结果表明,与空白组相比,植物组的O1、O2和N1类等化合物相对丰度都发生了显著变化,石油污染物得到一定程度的生物降解。     

石油污染土壤生物修复菌Z1a-B的分离鉴定与调控效应研究

从山东东营胜利油田附近的石油污染土壤中分离筛选得到一株高效石油降解菌Z1a-B,通过菌落形态及显微镜个体形态观察对其初步鉴定到属,并采用气相色谱/质谱(GC/MS)法分析了Z1a-B的石油降解性能,采用投加石油降解菌、调节土壤N、P含量和优化环境因素等措施,进行了为期60d的石油污染土壤室外自然堆制生物修复实验。结果表明,Z1a-B为链霉菌属白孢类群,其摇瓶培养的石油降解率为66.4%;Z1a-B有着很宽的烷烃降解谱;N、P最佳的添加量组合为KNO32.50g/kg、K2HPO40.35g/kg,即N/P(质量比)为5.57:1.00,此时的石油降解率达63.5%,土壤脱氢酶活性达最高值,为2.99μL/g;石油降解的最佳环境条件为:将石油质量分数为3.3%的100g土样调节pH至8.5后,装入容积为300mL的锥形瓶中灭菌,再接种孢子密度为2.7×108个/mL的菌剂5.5mL,于28℃下进行生物降解,在此条件下的石油降解率可达76.5%;土壤脱氢酶活性的测定结果可以作为检验石油污染土壤生物修复效果的重要指示指标之一;室外自然堆制生物修复实验中,添加菌剂、锯末、秸秆以及N、P后,石油降解率可达69.9%,总体来说,室外自然堆制生物修复是一种投资少、见效快、治理效果较好的石油污染土壤治理方法。     

红三叶草根际区石油降解菌的筛选及降解性能

从石油污染的土壤红三叶草（nifoliumrepensLinn）根际修复区中分离筛选得到4株以原油作为惟一碳源和能源进行生长繁殖的高效石油降解菌。通过菌落形态、显微镜个体形态观察、生理生化鉴定以及菌株16SrDNA序列分析，初步鉴定4株优势降解菌分别为动性杆菌、藤黄微球菌、蜡状芽孢杆菌和短小芽孢杆菌。采用气相色谱／质谱（GC／MS）法分析4株混合菌对石油烃的降解性能。结果表明：在摇床培养条件下，混合菌54d对总石油烃的生物降解率达到90．50％，较对照高67．72％。随着生物降解时间的延长，石油组分中的正构烷烃、异构烷烃及环烷烃相对总量均呈减小趋势，而芳香烃和其他醇类、醛和酸类的相对含量则有所增加。     

华北某油田石油降解菌的筛选及降解特性

以石油为唯一碳源,从华北某油田污染土壤中筛选出石油降解菌12株。其中5株菌有较强降解能力,分别编号为z-3、z-6、z-7、z-8和z-b,在30℃,160 r/min摇床培养10 d后,菌株对石油降解率分别为63.8%、34.2%、44.8%、50.5%和42.3%。通过生理生化和分子生物学鉴定,确定这5株菌分别属于假单胞菌属(Pseudomonas)、根瘤菌属(Rhizobium)、假黄单胞菌属(Pseudoxanthomonas)、假单胞菌属(Pseudomonas)、鞘氨醇单胞菌属(Sphingomonas)。通过对这5株菌生物量、脱氢酶及邻苯二酚2,3-双加氧酶活性的监测,发现生物量及酶活性与石油降解能力具有直接关系。最后,对5株菌的生长条件进行优化,其中z-3、z-6、z-7和z-b菌株对石油降解率较优化前提高10%左右。     

Nonionic surfactant enhanced biodegradation of m-xylene by mixed bacteria and its application in biotrickling filter

In this study, m-xylene biodegradation was examined in bacteria-water mixed solution and biotrickling filter (BTF) systems amended with the nonionic surfactant Tween 80. The mixed bacteria were obtained from the activated sludge of a coking plant through a multisubstrate acclimatization process. High-throughput sequencing analysis revealed that Rhodanobacter sp. was the dominant species among the mixed bacteria. In the bacteria-water mixed solution, the bacterial density increased with increasing Tween 80 concentration. Hence, Tween 80 could be utilized as substrate by the mixed bacteria. Tween 80, with concentrations of 50–100 mg L^?1, could enhance the bioavailability of m-xylene and consequently improve the degradation efficiency of m-xylene. However, further increasing the initial concentration of Tween 80 would decrease the degradation efficiency of m-xylene. At concentrations exceeding 100 mg L^?1, Tween 80 was preferentially degraded by the mixed bacteria over m-xylene. In BTF systems, when the m-xylene inlet concentration was 1200 mg m^?3 and the empty bed residence time was 20 sec, the removal efficiency and elimination capacity of BTF1 with Tween 80 addition were at most 20% and 24% higher than those of BTF2 without Tween 80 addition. Overall, the integrated application of the mixed bacteria and surfactant was demonstrated to be a highly effective strategy for m-xylene waste gas treatment.

Implications: The integrated application of mixed bacteria and surfactant was demonstrated to be a promising approach for the highly efficient removal of m-xylene. Surfactant can activate mixed bacteria to degrade m-xylene by increasing its bioavailability. Besides, surfactant can be utilized as carbon source by the mixed bacteria so that the growth of mixed bacteria can be promoted. It is expected that the integrated application of both technologies will become more common in future chemical industry.     

Preparation of petroleum-degrading bacterial agent and its application in remediation of contaminated soil in Shengli Oil Field,China

Two petroleum-degrading strains were screened from oil fields and denoted as SWH-1 (Bacillus subtilis) and SWH-2 (Sphingobacterium multivorum), which were used to ferment and prepare bacterial agent to remediate petroleum-contaminated sites in Shengli Oil Field in China. The optimal liquid fermentation medium and conditions were MgSO₄·7H₂O (0.5 %), NaCl (0.5 %), soybean dregs (3 %), pH 7.0, culturing at 30 °C, and 220 r/min for 16 h. Peat was chosen as the bacterial carrier due to its ability of keeping microbial activity. Mixed fermented liquid was added into peat (1:2) and air-dried, and the bacterial agent was obtained. It was applied to the petroleum-contaminated soil, which was irrigated, tilled, and fertilized. The removal rate reached 67.7 % after 2 months of remediation. During remediation, the quantity of indigenous bacteria varied a lot, while the inoculated bacteria remained stable; the dehydrogenase activity was at high levels and then decreased. Indigenous microorganisms, inoculated bacterial agent, nutrients, water, and soil permeability all played important roles. The study prepared an environment-friendly bacterial agent and established a set of bioremediation technique, which provided further insights into integration of fermentation engineering and soil remediation engineering.     

Anaerobic degradation of five polycyclic aromatic hydrocarbons from river sediment in Taiwan

This study investigated the anaerobic degradation of five polycyclic aromatic hydrocarbons (PAHs) from Erren River sediment in southern Taiwan. The degradation rates of PAH were in the order: acenaphthene > fluorene > phenanthrene > anthracene > pyrene. The degradation rate was enhanced when the five compounds were present simultaneously in river sediment. Comparison of the PAH degradation rates under three reducing conditions showed the following order: sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The addition of electron donors (acetate, lactate and pyruvate) enhanced PAH degradation under methanogenic and sulfate-reducing conditions. However, the addition of acetate, lactate or pyruvate inhibited PAH degradation under nitrate-reducing conditions. The addition of heavy metals, nonylphenol and phthalate esters (PAEs) inhibited PAH degradation. Our results show that sulfate-reducing bacteria, methanogen and eubacteria are involved in the degradation of PAH; sulfate-reducing bacteria constitute a major microbial component in PAH degradation. Of the microorganism strains isolated from the sediment samples, we found that strain ER9 expressed the greatest biodegrading ability.     

Anaerobic degradation of five polycyclic aromatic hydrocarbons from river sediment in Taiwan

This study investigated the anaerobic degradation of five polycyclic aromatic hydrocarbons (PAHs) from Erren River sediment in southern Taiwan. The degradation rates of PAH were in the order: acenaphthene > fluorene > phenanthrene > anthracene > pyrene. The degradation rate was enhanced when the five compounds were present simultaneously in river sediment. Comparison of the PAH degradation rates under three reducing conditions showed the following order: sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The addition of electron donors (acetate, lactate and pyruvate) enhanced PAH degradation under methanogenic and sulfate-reducing conditions. However, the addition of acetate, lactate or pyruvate inhibited PAH degradation under nitrate-reducing conditions. The addition of heavy metals, nonylphenol and phthalate esters (PAEs) inhibited PAH degradation. Our results show that sulfate-reducing bacteria, methanogen and eubacteria are involved in the degradation of PAH; sulfate-reducing bacteria constitute a major microbial component in PAH degradation. Of the microorganism strains isolated from the sediment samples, we found that strain ER9 expressed the greatest biodegrading ability.     

Effect of UV radiation and temperature on mineralization and volatilization of coumaphos in water

This study was undertaken to determine the dissipation and degradation of coumaphos [O-(3-chloro-4-methyl-2-oxo-2H-1-benzopyran-7-yl) O,O-diethyl phosphorothioate] under different sunlight conditions and at different temperatures. The effect of the ultra violet (UV) component of solar radiation was also studied using quartz tubes in addition to other radiation in the visible range using glass tubes and the results were compared with those obtained under the dark light conditions. Water suspensions of coumaphos were incubated at three temperatures viz. 22°C, 37°C and 53°C in closed systems to study the effect of temperature. Volatilization, mineralization and degradation of coumaphos increased with an increase in temperature and exposure to solar radiation, particularly under the UV component of the solar radiation. Major loss of the pesticide occurred through volatilization. The optimum temperature for the degradation of coumaphos was found to be at 37°C. The data obtained from the mineralization and degradation studies indicated that 53°C crosses the biological range for suitable growth of microorganism. UV radiation exposure along with maintaining temperature at 37°C may prove useful in the dissipation and/or degradation of coumaphos prior to its disposal as waste from cattle dipping vats.     

Bioremediation of oil-contaminated soil using Candida catenulata and food waste

Even though petroleum-degrading microorganisms are widely distributed in soil and water, they may not be present in sufficient numbers to achieve contaminant remediation. In such cases, it may be useful to inoculate the polluted area with highly effective petroleum-degrading microbial strains to augment the exiting ones. In order to identify a microbial strain for bioaugmentation of oil-contaminated soil, we isolated a microbial strain with high emulsification and petroleum hydrocarbon degradation efficiency of diesel fuel in culture. The efficacy of the isolated microbial strain, identified as Candida catenulata CM1, was further evaluated during composting of a mixture containing 23% food waste and 77% diesel-contaminated soil including 2% (w/w) diesel. After 13 days of composting, 84% of the initial petroleum hydrocarbon was degraded in composting mixes containing a powdered form of CM1 (CM1-solid), compared with 48% of removal ratio in control reactor without inoculum. This finding suggests that CM1 is a viable microbial strain for bioremediation of oil-contaminated soil with food waste through composting processes.     

鼠李糖脂对微生物菌剂降解石油的影响

以石油烃降解微生物菌剂和铜绿假单胞菌株A6为对象，考察不同浓度鼠李糖脂对菌剂细胞表面疏水性、原油降解性能和微生物生长的影响，采用GC-MS分析石油中正构烷烃组分的降解情况。结果表明，低浓度鼠李糖脂就可提高菌剂细胞的表面疏水性和原油降解效果。以250mg／L添加组最明显，第7天疏水性达最高，为58．6％，比对照组提高约26．2％；降解第15天原油降解率达71．6％，分别比对照组和TW20组提高16％和13．3％。GC-MS结果显示鼠李糖脂对高碳数烷烃的降解作用大于低碳数烷烃，正二十三烷和正三十三烷的降解率分别较对照提高了21．5％和33．7％。菌剂对奇数碳烷烃的降解效果优于偶数碳烷烃。鼠李糖脂分别使菌剂中细菌、放线菌和霉菌的最大生物量提高了5．7、2．4和1．8倍。鼠李糖脂对微生物细胞疏水性和生物量的提高与石油降解效果正相关。     

菌源对多环芳烃降解菌的筛选及降解性能的影响

高效降解菌的筛选对利用生物修复技术有效去除环境中的多环芳烃具有重要意义。分别以石油污染土壤和焦化废水活性污泥为菌源,分离出芘降解菌和混合PAHs(菲、荧蒽和芘)降解菌共14株并对其降解性能进行对比研究。结果表明,筛选得到的菌株分别属于9个菌属,其中2种菌源共有的菌属为Mycobacterium sp.、Ralstonia sp.和Shinella sp.。芘和PAHs的高效降解菌(CP16和CM32)均属于分支杆菌属(Mycobacterium),来源于焦化废水活性污泥;菌株CP16对芘(50mg/L)的7 d降解率为74.99%,CM32对PAHs(菲50 mg/L、荧蒽和芘各10 mg/L)的7 d降解率为100%。因此,以焦化废水活性污泥为菌源更有利于获得高效的多环芳烃降解菌。     

Assessment of a process to degrade metal working fluids using Pseudomonas stutzeri CECT 930 and indigenous microbial consortia

The development of a novel biological process to treat metal working fluids (MWFs)-containing effluents at bioreactor scale was pursued in this work. The bacteria Pseudomonas stutzeri CECT 930 was investigated for the first time as an alternative agent for MWF degradation. An adequate medium design and mixing and aeration system, as well as an appropriate microorganism proved to be crucial for reaching high levels of degradation by P. stutzeri and by an indigenous consortium (about 70% and 50% of reduction in total petroleum hydrocarbon content in less than 2 wk, respectively). Additionally, as there is no information in literature trying to kinetically characterize an MWF-polluted effluent degradation process, all the experimental data were fitted to logistic and Luedeking and Piret models, that allowed to elucidate the growth-associated character of the biodegradation process.