寡养单胞菌降解石油污染土壤中的甲苯

石油污染土壤治理一直是环境污染控制技术的研究重点,微生物修复石油污染是一种高效、低廉的处理技术。从石油污染的土壤中筛选出了1株专一性降解甲苯的菌株寡养单胞菌(Stenotrophomonas sp.MJ-1),研究了甲苯浓度和温度对降解的影响,并对动力学和热力学进行了探讨。结果表明:菌株Stenotrophomonas sp.MJ-1在温度为30℃,甲苯浓度为5 mg/L时降解效果最好,最大去除率可达90%。菌株降解污染物的过程均符合一级动力学模型。热力学研究表明菌株Stenotrophomonas sp.MJ-1对甲苯降解反应活化能E_a为17.27 k J/mol。     

甲硫醇降解菌群筛选及其降解特性研究

从处理甲硫醚(DMS)和丙硫醇(PT)混合废气的生物滴滤塔中富集出一组能够有效降解甲硫醇(MT)的混合菌群,并对其特性进行了系列研究.结果表明,该混合菌群能有效降解MT,菌群较为适宜的生长和降解条件为30℃、p H=7.0,在该条件下能将初始浓度为20 mg·L~(-1)的MT在70 h内降解完全.添加酵母膏(YE)后,MT降解速率进一步提高,降解所需时间缩短10 h.利用高通量测序技术分析混合菌群的群落结构,发现其中优势菌属为Pseudomonas sp.、Thiobacillus sp.和Acinetobacter sp.,所占比例分别为33.78%、21.91%和17.01%.中间产物检测结果表明,混合菌群降解MT的过程中产生了甲醛、H_2S、二甲基二硫醚(DMDS)等物质,推断MT的降解途径可能有如下2条:(1)MT在MT氧化酶作用下形成甲醛和H2S,随后氧化为SO_4~(2-);(2)MT依次转化为DMDS、DMS、二甲基亚砜(DMSO)和二甲基砜(DMSO_2),最后经甲基磺酸(MSA)可生成SO_4~(2-).     

降解石油丝状真菌的筛选及其降解特性

从大庆石油污染土壤中分离得到14株石油降解丝状真菌,用以筛选可高效降解石油的菌群. 经过ITS(转录间隔区序列)分析,其中9株属于镰孢霉属(Fusarium sp.),2株属于黄白生丛赤壳菌属(Bionectria sp.),另外3株分别属于葡萄穗酶菌属(Stachybotrys sp.)、曲霉属(Aspergillus sp.)和雅致放射毛霉属(Actinomucor sp.);在固体培养基中各菌株的生长速率差异显著(P<0.01). 不同菌株能够特异降解不同的石油组分. 将筛选的优势菌株组成2个菌群,菌群1由菌株3、D2、D3和D52组成,菌群2由菌株3、6、D2和D3组成,分别考察单菌和2个菌群对原油的降解效果,结果表明:菌株原油降解试验30d后,单菌菌株D52对原油去除率最大,为64.25%;菌群1和菌群2对原油的去除率较高,分别达到74.55%和72.64%,可以考虑用于污油生物修复治理的工程菌群开发研究.      

石油降解低温细菌的筛选及降解特性的表征

从石油污染土壤中,通过低温富集,筛选并鉴定得到7株低温石油降解细菌。基于菌株降解石油组分特性,构建6组低温石油降解菌群,利用5 L发酵罐,并通过尾气分析仪在线监测菌群石油降解过程中的CO₂产生和O₂消耗变化,评价菌群的石油降解能力。由Arthrobacter sp. JLH 001,Acinetobacter baumannii JLH 002,Pseudomonas fragi JLH 003和Arthrobacter sp. JLH 006组成的菌群降解石油效果最佳,48 h后CO₂的产生值和O₂的消耗值达到最高,在15 ℃时、72 h后能完全降解1%的石油,并且在25 ℃时降解速度显著增强。结果表明:石油污染土壤的原位生物修复可通过低温石油降解菌群的添加实现高效及快速修复。     

高效复合菌对多菌灵的生物降解

将多菌灵降解菌Alcaligenes sp.和Rhodococcua sp.(编号为A和R)按不同比例进行复配,并采用三波长校正法和HPLC法测定不同复配降解体系中多菌灵的残留量,比较了纯培养和复合菌群对多菌灵的降解效果,最后对高效复合菌的降解条件进行了优化.试验结果表明,得到的高效降解复合菌群AR5(A∶R复配比例1...     

MBR中微生物群落结构的演变与分析

为了揭示膜.生物反应器中微生物群落结构多样性的演变过程,通过细胞裂解法直接提取不同时期污泥中的基因组DNA,利用基于16SrDNA的PCR-DGGE技术获得了微生物群落的DNA特征指纹图谱,并对条带进行了统计分析和切胶测序,使用序列数据进行了同源性分析并建立了系统发育树.DGGE分析表明,在反应器运行前17d内污泥中微生物群落结构变化很大,与接种污泥的相似性系数下降到了29.2%,从而说明MBR中处理工艺和进水水质的改变导致微生物群落结构多样性降低.在试验过程中,Pscudomonas和Aeromonas hydrophila等种群一直保持着较为稳定的优势地位,也有原始种群如Bacillus sp.的消亡和以Enterococcus faecalis、Comamonas sp.、Fusobacterium sp.等为代表的次级种群的强化和演变.UPGMA聚类分析将DGGE图谱区分为3大类群并对应于各自的运行时期.测序结果表明,MBR中微生物菌群间进化距离较大,其中Proteobaeteria纲和Bacillus属细菌较多.在反应器运行后期演变为优势地位的菌群(如Comamonas sp.)加剧了膜污染物的产生和积累.     

质粒基因强化活性污泥系统对2,4–D的降解

以携带质粒pJP4[含编码2,4-二氯苯氧基乙酸(2,4-D)降解功能的基因片段]的基因工程菌Pseudomonas putida SM1443::gfp2x (pJP4::dsRed)为供体菌,通过半连续流实验研究了质粒基因强化对活性污泥系统的2,4-D的降解效应及菌群结构的影响.结果表明,以初始浓度约为320mg/L的2,4-D为唯一碳源,向活性污泥系统投加携带pJP4质粒的基因工程菌P. putida,运行初期,降解促进作用不明显;随着半连续流反应的进行,促进作用显著增强.与对照系统相比,降解速率之差最高为6.67mg/(L·h).从基因强化系统中筛选到1株占有优势的接合子,经鉴定为Alcaligenes sp.::pJP4. Alcaligenes sp.本身不具备降解2,4-D的能力,获得质粒pJP4后,对2,4-D降解能力大幅度提高,与野生型2,4-D高效降解菌Bacillus sp.相当. PCR-DGGE分析表明,在受到2,4-D冲击条件下基因强化的活性污泥系统较对照系统保持了相对更加稳定的菌群结构.     

1株异养脱氮菌降解壬基酚聚氧乙烯醚的研究

研究了异养脱氮菌Bacillus sp.LY降解壬基酚聚氧乙烯醚(NPEOs)的性能.结果表明,该菌株具有较强地降解NPEOs的能力,且在实现NPEOs降解去除的同时表现出一定的异养脱氮性能.降解14d后, Bacillus sp.LY对NPEOs去除率达95.6%,对体系中的总氮去除率为43.9%.该菌株对NPEOs的降解去除符合一级动力学特征,其降解速率常数为0.224d-1.该菌株通过无氧化过程的乙氧基链的逐渐缩短的途径降解去除NPEOs,可避免产生危害性更大的NPEOs的羧酸化产物(NPECs).在分别以氨态氮(NH4C1)、硝态氮(NaNO3)和亚硝态氮(NaNO2)3种不同氮素为氮源的条件下,菌株对NPEOs均具有一定的降解效果,其中以氨态氮为氮源时菌株对NPEOs降解效果最好.研究结果可为消除壬基酚聚氧乙烯醚与氮素复合污染提供理论依据.     

苯系化合物在硝酸盐还原条件下的生物降解性能

运用驯化的反硝化混合菌群进行了苯系化合物(BTEX)的厌氧降解试验.结果表明,混合菌群能够在反硝化条件下有效降解苯、甲苯、乙苯、邻二甲苯、间二甲苯和对二甲苯.BTEX的降解规律符合底物抑制的Monod模型,当初始浓度小于50mg·L^-1时,6种受试基质的厌氧降解速率顺序为:甲苯>乙苯>间二甲苯>邻二甲苯>对二甲苯>苯.整个试验过程中NO₃^-的消耗与苯、甲苯、乙苯、邻二甲苯、间二甲苯及对二甲苯生物降解之间的摩尔比分别为:9.47,9.26,1     

碱性缺氧环境下地下水中苯和甲苯的生物降解

在缺氧环境下,不额外加入电子受体和营养盐,从长期受原油污染的包气带介质中分离、培养驯化得到了降解苯或甲苯的3种优势菌群:B-bacteria、T-bacteria和M-bacteria,采用批试验方法研究了高pH环境下3种菌群降解苯和甲苯的速率。结果表明:苯和甲苯的降解符合零级反应动力学,速率常数在0.22～0.68 mg/(L.d)。初始pH从8.7升高到9.6和10.6时,B-bacteria降解苯的速率降低都在10%以内;T-bacteria降解甲苯的速率降低率从pH9.6时的16.22%剧增到pH10.6时的41.23%;而M-bacteria降解苯和甲苯的速率降低从pH9.6时的30%左右增到pH10.6时的45%左右。高pH环境下微生物仍能完全降解苯和甲苯。故设计化学-生物连续反应格栅治理该类污染羽时,在两个单元中间可不构筑pH调节缓冲单元。     

水解酸化-缺氧生物法处理油田废水的机理

采用水解酸化-缺氧生物法对经物化预处理的油田废水进行试验研究.当进水COD为190～230mg/L,水解酸化段和缺氧段停留时间分别为10,48h时,出水COD为75～83mg/L.运用GC/MS分析油田废水有机污染物在工艺流程中相对组分变化的规律,表明水解酸化和缺氧法处理油田废水时有协同作用,可有效降解废水中酚类化合物、酮类化合物、芳烃和BTEX.运用PCR-DGGE技术,考察不同生物反应器内微生物种群及其分布特征,初步确定水解酸化和缺氧反应器内的优势菌种.     

Effcient degradation of lube oil by a mixed bacterial consortium

A laboratory study was performed to assess the biodegradation of lube oil in bio-reactor with 304# stainless steel as a biofilm carrier. Among 164 oil degrading bacterial cultures isolated from oil contaminated soil samples, Commaonas acidovorans Px1, Bacillus sp. Px2, Pseudomonas sp. Px3 were selected to prepare a mixed consortium for the study based on the e ciency of lube oil utilization. The percentage of oil degraded by the mixed bacterial consortium decreased slightly from 99% to 97.2% as the concentration of lube oil was increased from 2000 to 10,000 mg/L. The degradation of TDOC (total dissolved organic carbon) showed a similar tendency compared with lube oil removal, which indicated that the intermediates in degradation process hardly accumulated. Selected mixed bacterial consortium showed their edge compared to activated sludge. Scanning electron microscopy (SEM) photos showed that biofilms on stainless steel were robust and with a dimensional framework constructed by EPS (extracellular polymeric substances), which could promote the biodegradation of hydrocarbons. The increase of biofilm followed first-order kinetics with rate of 0.216 g glucose/(cm2 day) in logarithm phase. With analysis of Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS) combined with removal of lube oil and TDOC, mixed bacterial consortium could degrade benzene and its derivatives, aromatic ring organic matters with a percentage over 97%.     

Efficient degradation of lube off by a mixed bacterial consortium

A laboratory study was performed to assess the biodegradation of lube oil in bio-reactor with 304# stainless steel as a biofilm carrier.Among 164 oil degrading bacterial cultures isolated from oil contaminated soil samples, Commaonas acidovorans Pxl, Bacillus sp.Px2, Pseudomonas sp. Px3 were selected to prepare a mixed consortium for the study based on the efficiency of lube oil utilization.The percentage of oil degraded by the mixed bacterial consortium decreased slightly from 99% to 97.2% as the concentration of lube oil was increased from 2000 to 10,000 mg/L. The degradation of TDOC (total dissolved organic carbon) showed a similar tendency compared with lube oil removal, which indicated that the intermediates in degradation process hardly accumulated. Selected mixed bacterial consortium showed their edge compared to activated sludge. Scanning electron microscopy (SEM) photos showed that biofilms on stainless steel were robust and with a dimensional framework constructed by EPS (extracellular polymeric substances),which could promote the biodegradation of hydrocarbons. The increase of biofilm followed first-order kinetics with rate of 0.216 μg glucose/(cm~2·day) in logarithm phase. With analysis of Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS) combined with removal of lube oil and TDOC, mixed bacterial consortium could degrade benzene and its derivatives, aromatic ring organic matters with a percentage over 97%.     

Isolation and characterization of gasoline-degrading bacteria from gas station leaking-contaminated soils

The effects of culture conditions in vitro and biosurfactant detection were studied on bacterial strains capable of degrading gasoline from contaminated soils near gas station. The main results were summarized as follows. Three bacteria （strains Q10, Q14 and Q18） that were considered as efficiently degrading strains were isolated and identified as Pseudomonas sp., Flavobaeterium sp. and Rhodococcus sp., respectively. The optimal growth conditions of three bacteria including pH, temperature and the concentration of gasoline were similar. The reduction in surface tension was observed with all the three bacteria, indicating the production of biosurfactant compounds. The value of surface tension reduced by the three strains Q10, Q14 and Q18 was 32.6 mN.m, 12.4 mN. m and 21.9 mN.m, respectively. Strain Q10 could be considered as a potential biosurfactant producer. Gasoline, diesel oil, benzene, toluene, ethylbenzene and xylene （BTEX） could easily be degraded by the three isolates. The consortium was more effective than the individual cultures in degrading added gasoline, diesel oil, and BTEX. These results indicate that these strains have great potential for in situ remediation of soils contaminated by gas station leaking.     

嗜盐菌群对菲的降解及萘双加氧酶基因的表达规律

从胜利油田富集了一个能够降解多环芳烃的嗜盐菌群,通过克隆文库技术解析了菌群萘双加氧酶(ndo)基因的多样性.结果表明,该嗜盐菌群有6种ndo基因型,其中3种主要基因型(占总克隆子92.7%)与经典nah-like基因的相似度为89%.采用real-time RT-PCR等技术分析了不同盐度下菲降解过程中ndo基因的表达量、降解速率、生长曲线以及菲的生物可利用度,探索了菌群对菲的降解及ndo基因的表达规律.结果表明,当盐度由10%升高到20%,菌群完全降解100mg/L菲的时间从6d延长到9d,菌群生长的迟滞期由1d延长为3d.溶解菲的浓度在菌群生长过程中呈增加的趋势.Ndo基因的表达量在降解过程中呈先降低后随溶解菲浓度升高而升高的趋势,表明高浓度菲在初始阶段可能抑制ndo基因的表达.单因素方差分析(n=3, P>0.05)表明溶解菲浓度在10%和20%盐度下没有显著差异,表明一定范围的盐度不会影响菌群降解过程中菲的溶解度.菌群ndo基因的相对表达量在10%盐度下较在20%盐度下高,说明盐度对嗜盐菌群功能基因的表达具有抑制作用.     

Characterization of a salt-tolerant bacterium Bacillus sp. from a membrane bioreactor for saline wastewater treatment

High salt concentrations can cause plasmolysis and loss of activity of cells, but the salt-torlerant bacterium can endure the high salt concentrations in wastewater. In this research 7 salt-torlerant bacteria, which could survive in dry powder products and could degrade organic contaminants in saline wastewater, were isolated from a membrane bioreactor. The strain NY6 which showed the fastest growth rate, best property for organic matter degradation and could survive in dry powder more than 3 months was selected and characterized. It was classified as Bacillus aerius based on the analysis of the morphological and physiological properties as well as the 16 S rRNA sequence and Neigh borjoining tree. The strain NY6 could survive in the salinity up to 6% and the optimal growth salinity is2%; it belongs to a slightly halophilic bacterium. The capability of its dry powder products for COD removal was 800 mg COD/(g·day) in synthesized saline wastewater with salinity of 2%. According to salt-tolerant mechanism research, when the salinity was below 2%, the stain NY6 absorbed K+and Na+to maintain osmotic equilibrium, and when the salinity was above 2%, the NY6 kept its life by producing a large amount of spores.     

生物油降解菌的分离鉴定及其在土壤中的降解特性

从造纸厂内废水处理池中采取活性污泥,筛选出一株能高效降解生物油的菌株EL5.对该菌株在土壤中对生物油降解能力进行了初步研究;建立了基于CO₂生成量的生物降解测定方法,并运用该方法,对生物油生物降解性能进行了评定.结果表明:①通过富集培养方法分离得到菌株EL5,根据形态特征以及生理化特性分析,将EL5鉴定为杂色曲霉菌(Aspergill versicoir).②菌株EL5对5种常见抗生素敏感.③菌株EL5在降解生物油的过程中,生物降解率与温度呈正相关关系.考虑到实际应用情况,选择30 ℃为适宜降解温度.pH为中性条件下对生物油的降解较为有利.随着w(生物油)的增加生物降解率有一定的下降,但相同条件下,加菌后30 ℃下10 d对w(生物油)为1 200 mg/kg的土壤的生物降解率达40%,而未加菌的生物降解率只有6%.④菌株EL5还可以以苯、对氯苯酚、甲苯和二甲苯等芳香类化合物作为唯一碳源生长繁殖.      

大学校园室内BTEX浓度水平变化规律及健康风险

采用Tenax-TA吸附/热脱附-气相色谱法(TD-GC)对大学校园室内外空气中5种苯系物(BTEX,苯、甲苯、乙苯、间/对-二甲苯和邻二甲苯)的平均浓度进行了检测。检测结果显示,5种苯系物的平均浓度均低于国家标准值。被测空气的苯系物中甲苯所占比例最大,为27.9%～32.0%。室内BTEX浓度稍高于室外,多数采样点的室内浓度与室外浓度比值(CI/CO)大于1.0。通风可有效降低空气中苯系物浓度。大学校园室内空气中的苯对学生的致癌风险为3.67×10-7～1.09×10-6。     

硝酸盐和甲烷对覆土中苯系物厌氧氧化的影响

苯系物是填埋场填埋气中恶臭有机气体的重要成分之一,填埋气中的CH4则是重要的温室气体.填埋覆土层中的微生物可以氧化CH4和苯系物,因此,强化微生物的氧化效能有助于削减和控制填埋气的污染.电子受体还原可耦合甲烷和某些有机物的厌氧氧化,从而去除甲烷和有机物.鉴此,本研究通过静态培养试验,分析了电子受体SO2-4共存条件下,NO-3和CH4共存对覆土中苯系物厌氧降解的影响.结果表明不外加NO-3时,苯系物抑制CH4的降解,加入NO-3后,苯系物共存反而有利于CH4的去除;单独添加NO-3或CH4都能促进填埋覆土中苯系物的去除;而同时添加NO-3和CH4能更好地促进苯系物的去除,甲苯、二甲苯和异丙苯的去除率最高可达65%、88%和82%,远高于不添加NO-3和CH4对照处理的53%、76%和31%;NO-3还原与CH4厌氧氧化耦合过程能同步促进苯系物的厌氧氧化.     

耐盐石油烃降解菌的筛选鉴定及其特性研究

为得到高效耐盐石油烃降解菌,从黄河三角洲石油污染盐渍化土壤中分离出39株细菌,经液体培养初筛和土壤培养复筛实验,得到1株高效耐盐石油烃降解菌BM38.通过形态特征、生理生化特征和16S rDNA序列分析,确定该菌为恶臭假单胞菌（Pseudomonas putida）.通过液体培养实验,研究了BM38的耐盐和产生物表面活性剂特性以及对不同烃的利用能力.结果表明,在含0.5%～6.0%NaCl液体培养基中BM38生长良好,属中度耐盐菌.在高盐环境下BM38具有较强的分解石油烃能力,其中在含1.0%NaCl液体培养基中,降解7 d后,原油降解率达到73.5%;在含盐量0.22%和0.61%土壤中添加BM38,降解40 d后,土壤总石油烃降解率达到40%以上.BM38能产生一种生物乳化剂,盐浓度对这种乳化剂的乳化能力影响较大,当NaCl浓度增加到1.0%,乳化值（EI24）开始迅速降低,但在NaCl浓度为2.0%时,EI24仍达到61.0%.BM38能够利用环己烷、甲苯、异辛烷、菲和正十六烷为唯一碳源生长,其中对正构烷烃和芳烃具有较强的利用能力.