Evaluation of biostimulation and Tween 80 addition for the bioremediation of long-term DDT-contaminated soil

The bioremediation of a long-term contaminated soil through biostimulation and surfactant addition was evaluated. The concentrations of 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane(DDT) and its metabolites 1,1-dichloro-2,2-bis(4-chlorophenyl) ethane(DDD) and1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene(DDE) were monitored during an 8-week remediation process. Physicochemical characterization of the treated soil was performed before and after the bioremediation process. The isolation and identification of predominant microorganisms during the remediation process were also carried out. The efficiency of detoxification was evaluated after each bioremediation protocol. Humidity and p H and the heterotrophic microorganism count were monitored weekly. The DDT concentration was reduced by 79% after 8 weeks via biostimulation with surfactant addition(B + S) and 94.3%via biostimulation alone(B). Likewise, the concentrations of the metabolites DDE and DDD were reduced to levels below the quantification limits. The microorganisms isolated during bioremediation were identified as Bacillus thuringiensis, Flavobacterium sp., Cuprivadius sp.,Variovorax soli, Phenylobacterium sp. and Lysobacter sp., among others. Analysis with scanning electron microscopy(SEM) allowed visualization of the colonization patterns of soil particles. The toxicity of the soil before and after bioremediation was evaluated using Vibrio fischeri as a bioluminescent sensor. A decrease in the toxic potential of the soil was verified by the increase of the concentration/effect relationship EC50 to 26.9% and 27.2% for B + S and B, respectively, compared to 0.4% obtained for the soil before treatment and 2.5%by natural attenuation after 8 weeks of treatment.     

Evaluation of biostimulation and Tween 80 addition for the bioremediation of long-term DDT-contaminated soil

The bioremediation of a long-term contaminated soil through biostimulation and surfactant addition was evaluated. The concentrations of 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) and its metabolites 1,1-dichloro-2,2-bis(4-chlorophenyl) ethane (DDD) and 1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene (DDE) were monitored during an 8-week remediation process. Physicochemical characterization of the treated soil was performed before and after the bioremediation process. The isolation and identification of predominant microorganisms during the remediation process were also carried out. The efficiency of detoxification was evaluated after each bioremediation protocol. Humidity and pH and the heterotrophic microorganism count were monitored weekly. The DDT concentration was reduced by 79% after 8 weeks via biostimulation with surfactant addition (B + S) and 94.3% via biostimulation alone (B). Likewise, the concentrations of the metabolites DDE and DDD were reduced to levels below the quantification limits. The microorganisms isolated during bioremediation were identified as Bacillus thuringiensis, Flavobacterium sp., Cuprivadius sp., Variovorax soli, Phenylobacterium sp. and Lysobacter sp., among others. Analysis with scanning electron microscopy (SEM) allowed visualization of the colonization patterns of soil particles. The toxicity of the soil before and after bioremediation was evaluated using Vibrio fischeri as a bioluminescent sensor. A decrease in the toxic potential of the soil was verified by the increase of the concentration/effect relationship EC₅₀ to 26.9% and 27.2% for B + S and B, respectively, compared to 0.4% obtained for the soil before treatment and 2.5% by natural attenuation after 8 weeks of treatment.     

Accelerated anaerobic dechlorination of DDT in slurry with Hydragric Acrisols using citric acid and anthraquinone-2,6-disulfonate (AQDS)

The application of electron donor and electron shuttle substances has a vital influence on electron transfer,thus may affect the reductive dechlorination of 1,1,1-trichoro-2,2-bis(p-chlorophenyl)ethane(DDT) in anaerobic reaction systems.To evaluate the roles of citric acid and anthraquinone-2,6-disulfonate(AQDS) in accelerating the reductive dechlorination of DDT in Hydragric Acrisols that contain abundant iron oxide,a batch anaerobic incubation experiment was conducted in a slurry system with four treatments of(1) control,(2) citric acid,(3) AQDS,and(4) citric acid + AQDS.Results showed that DDT residues decreased by 78.93%–92.11% of the initial quantities after 20 days of incubation,and 1,1-dichloro-2,2-bis(4-chlorophenyl)-ethane(DDD) was the dominant metabolite.The application of citric acid accelerated DDT dechlorination slightly in the first 8 days,while the methanogenesis rate increased quickly,and then the acceleration effect improved after the 8th day while the methanogenesis rate decreased.The amendment by AQDS decreased the Eh value of the reaction system and accelerated microbial reduction of Fe(III) oxides to generate Fe(II),which was an efficient electron donor,thus enhancing the reductive dechlorination rate of DDT.The addition of citric acid + AQDS was most efficient in stimulating DDT dechlorination,but no significant interaction between citric acid and AQDS on DDT dechlorination was observed.The results will be of great significance for developing an efficient in situ remediation strategy for DDT-contaminated sites.     

Study of DDT and its derivatives DDD, DDE adsorption and degradation over Fe-SBA-15 at low temperature

Mesoporous SBA-15 with different Fe₂O₃ loading were synthesized by an in-situ coating progress for removals of dichlorodiphenyltrichloroethane (DDT) and its derivatives, i.e., 1,1-dichloro-2,2-bis-(p-chlorophenyl)ethane (DDD) and 1,1-dichloro-2,2-bis-(4-chloro -phenyl) ethane (DDE). The results from XRD (X-ray diffractometer), TEM (transmission electron microscopy) indicated that the iron could be well dispersed on SBA-15 within 6 wt.% Fe₂O₃ loading. Nitrogen adsorption-desorption tests indicated that the synthesized materials were characterized by ordered meso-structure, high surface area and large pore volume. DDTs were removed from aqueous media in 12-hr treatment and high removal efficiency of DDTs was achieved at over 94%. DDTs could be completely degraded at 350℃ under the existence of SBA-15 with 4 wt.% Fe₂O₃ loading. The final degradation products of DDT were dichlorobenzophenone (DCB) and bis-(4-chloro-phenyl) methane (DDM), suggesting a complete dechlorination from trichloromethyl.     

Application of sewage sludge and intermittent aeration strategy to the bioremediation of DDT-and HCH-contaminated soil

Adding organic amendments to stimulate the biodegradation of pesticides is a subject of ongoing interest. The effect of sewage sludge on the bioremediation of dichlorodiphenyltrichloroethane（DDT） and hexachlorocyclohexane（HCH） contaminated soil was investigated in bench scale experiments,and intermittent aeration strategy was also used in this study to form an anaerobic–aerobic cycle. Bioremediation of DDT and HCH was enhanced with the addition of sewage sludge and the intermittent aeration. The removal rates of HCH and DDT were raised by 16.8%–80.8% in 10 days. Sewage sludge increased the organic carbon content from 6.2 to218 g/kg,and it could also introduce efficient degradation microbes to soil,including Pseudomonas sp.,Bacillus sp. and Sphingomonas sp. The unaerated phase enhanced the anaerobic dechlorination of DDT and HCH,and anaerobic removal rates of β-HCH,o,p′-DDT and p,p′-DDT accounted for more than 50% of the total removal rates,but the content of α-HCH declined more in the aerobic phase.     

1株筛自柴油污染土壤的铜绿假单胞菌对萘的降解特性研究

从柴油污染土壤中筛选分离出1株萘降解菌HD-5,经16S r DNA序列分析鉴定为铜绿假单胞菌(Pseudomonas aeruginosa),对功能基因进行PCR扩增证实该菌株中含有萘双加氧酶基因nah.采用生物强化、生物刺激以及二者相结合的方式修复萘含量为0.5%的自配污染土壤,综合比较了在不同修复方式下土壤中萘的降解率,修复过程中土壤FDA水解酶活和脱氢酶活的变化,以及运用定量PCR的方法动态分析了总细菌基因拷贝数和nah基因拷贝数.结果表明,在生物强化(B)、生物刺激(S)以及生物强化与生物刺激相结合(BS)这3种修复方式下,31 d后萘去除率分别为71.94%、62.22%和83.14%,BS组在修复过程中土壤FDA水解酶活和脱氢酶活明显高于另外两组,31 d后BS组土壤中总细菌基因拷贝数和nah基因拷贝数分别增长了约2.67×1011g-1和8.67×108g-1.上述研究结果表明筛选得到的萘降解菌株在土壤中具有良好的定植特性,在生物刺激与该降解菌株的共同作用下,可以有效地实现土壤中萘降解,这对此类污染生物修复过程研究具有一定的指导意义.     

一株DDT降解菌的筛选及其降解特性

以DDT为目标污染物,通过筛选获得了一株效果稳定的DDT降解菌,并对其进行形态学观察,生理生化特性及16S rRNA测序鉴定.经鉴定,该菌株属于甲基菌属（Methylovorus）,命名为Methylovorus sp. XLL03.菌株在pH值为7,温度30℃,外加碳源浓度0.5%,初始DDT浓度20mg/L时生长量最大.在pH值为6,温度30℃下,外加碳源（葡萄糖）浓度0.1%,初始DDT浓度20mg/L时对DDT的降解率最大.在优化条件下,4d后菌株XLL03对DDT最高降解率可达50.4%.利用GC-MS对DDT的降解中间产物进行定性分析,初步推断在菌株XLL03中,DDT最初分别通过脱氯和脱氯化氢生成DDD和DDE,随后DDD和DDE进一步脱氯得到DDMU,最终DDMU开环后又经过一系列反应被彻底矿化.在DDT的代谢过程中,未发现代谢中间产物的积累,表明菌株XLL03在修复受DDT污染的水或土壤中具有一定的应用前景.     

滴滴涕在我国典型POPs污染场地中的空间分布研究

在多年生产滴滴涕(DDT)的企业厂区内及周围布设采样点,采取不同深度的土壤进行分析,研究了POPs污染场地中DDT的空间分布规律.结果表明,生产厂区污染严重,在土壤表层,生产车间周围DDT的含量高达104mg·kg-1以上,离车间边界300m远处仍能检测到DDT,污染扩散范围与风向呈现一定相关性;在土壤深层,浓度随深度增加迅速降低,含量梯度变化最大为0.2～3m,3m以下土壤中浓度较小,10m深处浓度最高不超过8.89mg·kg-1.DDT的降解产物DDD和DDE在各个采样点均能检出,其降解率与含水率没有相关性.不同类型土壤中DDT的降解率存在很大差异,降解程度为粉砂(或粉土)＞黏土(或粉质黏土)＞杂填土.深层土壤中降解为DDD较为明显.(DDD DDE)/DDT比值显示,土壤中DDT为新输入的.     

旱地和淹水土壤中DDT的降解及DDMU和DDMS的形成趋势

在水稻和黑麦草2种栽培条件下,研究了新施入土壤中的工业DDT及微量老化残留DDT的降解,以及二级降解物DDMS和DDMU的形成动态.结果表明,DDT在旱地土壤中降解缓慢,其一级降解物DDE和DDD检出量略有增加,二级降解物DDMU和DDMS的形成极为有限,到试验终止时(黑麦草生长128 d)二者检出量仅为20 ng·g^-1.DDT在淹水土壤中可迅速降解,84 d后DDT残留量仅为施入量的2％左右,维持在70～80 ng·g^-1的水平;随DDT的降解,其一级降解物DDD检出量增加,在第84 d达峰值,DDD检出量的峰值与DDT的谷值时间基本吻合;同时DDMS和DDMU检出量也明显增加,在84 d时达到峰值,分别为28和115 ng·g^-1.未发现不同土壤条件对微量老化残留DDT降解动态或降解物形成产生影响.因此,淹水条件极利于新施入DDT在土壤中的降解并形成DDMS,但不能通过改变土壤水分条件来促进老化残留DDT降解.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

湖南省土壤中有机氯农药的残留规律研究

2004年5月采集了湖南省120个土壤样品,采用AES萃取技术,用GC-MS方法测定了样品中的六氯苯、滴滴涕（DDTs）、氯丹、艾氏剂、狄氏剂、异狄氏剂、七氯和灭蚁灵.结果显示:六氯苯和DDTs的检出率为100%,氯丹和灭蚁灵的检出率较低,艾氏剂、狄氏剂、异狄氏剂和七氯均未检出,说明六氯苯和DDTs曾在湖南省广泛施用.w（有机氯农药）平均值为115.3 μg/kg,其中w（DDTs）占w（有机氯农药）的96.44%,在全部120个样点中有21.67%的土壤样品的w（DDT）/w（DDE+DDD）大于1,表明DDTs曾是湖南省用于农作物的主要杀虫剂,并且近期仍然有输入.不同使用功能的土壤中有机氯农药的残留量也不同,表现为旱地中的残留量高于水稻田,菜地中以辣椒地的残留量最高,茶场土壤中的残留量最低.      

湘江流域土壤中有机氯农药的残留规律

2004年5月采集了湘江流域51个样品,用AES萃取技术,使用GC-MS方法测定了样品中的六氯苯(HCB)、滴滴涕(DDTs)、氯丹、艾氏剂、狄氏剂、异狄氏剂、七氯和灭蚁灵等有机氯农药(OCPs). 结果表明:HCB和DDTs的检出率为100%,氯丹和灭蚁灵的检出率较低,艾氏剂、狄氏剂、异狄氏剂和七氯未检出,说明HCB和DDTs曾在湘江流域广泛施用. w(OCPs)平均值为145.49 μg/kg,其中w(DDTs)占w(OCPs)的90.9%,且有些土壤样品中w(DDT)/w(DDE+DDD)较大,说明DDTs曾作为湘江流域的主要杀虫剂施用过,并且近期仍然有输入. 农药残留量与农药的施用量成正比,农药施用量高的衡阳地区,其土壤中的农药残留量最高. 不同使用功能的土壤中有机氯农药的残留量不同,表现为旱地中的残留量高于水稻地,蔬菜地中以辣椒地的残留量最高.      

Combined remediation of DDT congeners and cadmium in soil by Sphingobacterium sp. D-6 and Sedum alfredii Hance

Combined pollution of 1,1,1-trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) and cadmium (Cd) in agricultural soils is of great concern because they present serious risk to food security and human health.In order to develop a cost-effective and safe method for the removal of DDTs and Cd in soil,combined remediation of DDTs and Cd in soil by Sphingobacterium sp.D-6 and the hyperaccumulator,Sedum alfredii Hance was investigated.After treatment for 210 days,the degradation half-lives of DDTs in soils treated by strain D-6 decreased by 8.1% to 68.0% compared with those in the controls.The inoculation of strain D-6 into soil decreased the uptake of DDTs by pak choi and S.alfredii.The shoots/roots ratios of S.alfredii for the Cd accumulation ranged from 12.32 to 21.75.The Cd concentration in soil decreased to 65.8%-71.8% for S.alfredii treatment and 14.1%-58.2% for S.alfredii and strain D-6 combined treatment,respectively,compared with that in the control.The population size of the DDTs-degrading strain,Simpson index (1/D) and soil respiratory rate decreased in the early stage of treatment and then gradually increased,ultimately recovering to or exceeding the initial level.The results indicated that synchronous incorporation of strain D-6 and S.alfredii into soil was found to significantly (p 0.05) enhance the degradation of DDTs in soil and the hyperaccumulation of Cd in S.alfredii.It was concluded that strain D-6 and S.alfredii could be used successfully to control DDTs and Cd in contaminated soil.     

土壤及果树中HCH和DDT残留及分布规律研究

采用索氏提取、弗罗里土固相微萃取小柱净化结合GC-ECD测定了苹果园土壤和果树样品中4种六六六(HCH)和4种滴滴涕(DDT)的残留量,并对果树的根、枝、叶中HCH和DDT的分布规律进行了研究。实验结果表明,土壤和果树样品中均有HCH和DDT检出。土壤样品中两类农药的含量分别在0.51～3.97ng/g和1.46～15.55ng/g之间,其中β-HCH是六六六残留的主体,滴滴涕类的含量以p,p'-DDT为最高,并且(DDE+DDD)/DDT<1表明近期果园中有新的DDT输入。果树样品中,HCH和DDT的残留量分别在0.22～2.01ng/g和0.38～6.08ng/g之间,都要低于相应土壤中有机氯农药的浓度。果树中总OCPs的含量在4.88～17.94ng/g之间,是土壤和大气中OCPs污染共同作用的结果。根部的OCPs的含量明显高于枝和叶,其分布规律为根>叶>枝,这主要取决于根、枝、叶各组织的结构以及农药的理化性质。     

小分子有机碳源对滴滴涕污染沉积物生物修复作用的基础研究

考察了辽河沉积物p,p'-DDT在无外加碳源和外加乳酸钠或丙酸钠情况下的缺氧生的降解行为，降解过程中体系的pH变化，并测定了p,p'-DDT的主要缺氧降解产物及其浓度变化，结果表现，p,p'-DDT的缺氧脱氯行为符合准一级反应动力学，无外加碳源，DDT的缺氧脱氯降解有近一个月的滞后期，降解速率为k（无碳源）＝0.0082d^-1，而外加碳源物质，大大缩短了DDT缺氧脱氯降解的滞后，降解速率分别为k(乳酸钠）＝0.0917d^-1,k(丙酸钠）＝0.1023d^-1，而且，p,p'-DDT的缺氧还原脱氯行为主要发生在体系中pH上升的阶段，p-p'-DDT缺氧生物降解的主要产物是p,p'-DDD，并有少量的p,p'-DDE产生；p,p'-DDD主要发生在体系中pH上升的阶段，p,p'-DDT缺氧生物降解的主要产物是p,p'-DDD，并有少量的p,p'-DDE产生，p,p'-DDD在后期有降解。     

府河和白洋淀沉积物中DDTs的分布特征和风险评估

利用气相色谱-质谱(GC-MS)检测了府河和白洋淀共计19个表层沉积物样品中的w(DDTs)及其分布特征. 结果表明,w(DDTs)为1.74～51.33 ng/g (以干质量计),平均值为11.01 ng/g,其分布特征呈现从府河到白洋淀逐渐递减的趋势. 与国内其他地区沉积物相比,府河和白洋淀地区DDTs的污染处于中等偏低的水平. 在所有沉积物样品中,p,p′-DDT所占比例较大,是最主要的异构体. 大部分样品中w(DDD)/w(DDE)大于1,而w(DDT)/w(DDE+DDD)小于1,说明研究区DDT处于厌氧环境条件,且没有新的DDT输入. 该地区DDTs主要来源于农田土壤侵蚀及工业废水排放. 潜在风险评估表明,府河和白洋淀的表层沉积物中DDTs对周围环境具有一定的影响,应引起相关部门的重视.      

基于高通量测序分析的生物修复石油污染土壤菌群结构变化

利用高通量测序技术对微生物修复石油污染土壤过程中的微生物群落结构变化进行研究.结果表明,经修复处理的土壤微生物群落结构及多样性发生明显变化.利用生物强化修复处理(BA)的土壤中,微生物丰富度与均匀度明显降低,土著菌群受到抑制,外加变形菌门(Proteobacteria)成为主要的优势菌门,相对丰度由修复前的37. 44%增加为87. 44%.假单胞菌属(Pseudomonas)成为土壤中的优势菌属,丰度由2. 99%增加为76. 37%;进行生物刺激修复处理的土壤(BS)菌群丰富度和均匀度与原污染土壤相比略有降低.菌群结构组成上,原优势菌门变形菌门(Proteobacteria)丰度由37. 44%降低为10. 90%,厚壁菌门(Firmicutes)丰度由9. 16%增加为35. 32%,属水平上,原优势菌属微小杆菌属(Exiguobacterium)和原小单胞菌属(Promicromonospora)丰度由8. 49%和18. 96%分别降低为2. 19%和14. 97%,诺卡氏菌属(Nocardioides)和芽孢杆菌属(Bacillus)丰度由5. 56%和0. 29%分别增加至28. 95%和22. 70%,成为主要优势菌属.生物强化修复处理引起土壤菌群多样性和结构发生明显变化,生物刺激修复处理可基本保持土壤菌群结构多样性不被破坏,土壤菌群结构的稳定有利于石油烃的生物降解.     

农药污染土壤中DDTs的电动力学迁移去除特性

为研究土壤中DDTs(滴滴涕)的电动迁移去除特性,以重庆市某废弃DDTs生产企业污染土壤为研究对象,采用GC-ECD分析了该土壤中DDTs的质量分数、电动迁移效果、电动去除率及试验过程中土壤理化性质改变等.结果表明:该污染场地土壤中w(p,p′-DDT)最高,w(o,p′-DDE)和w(p,p′-DDD)次之,三者分别占w(DDTs)的53.02%、23.46%和12.77%;DDTs迁移效果在192 h时基本稳定,并在3 V/cm电压梯度时迁移效果最佳,其中DDT和DDD向阳极迁移而DDE向阴极迁移,o,p′-DDE富集量最大,其c/c₀〔标准化浓度,c₀、c分别为电动试验前、后污染土壤中w(DDTs)〕为4.70;DDTs去除率在电动时间为192 h、电压梯度为3 V/cm时达到相对稳定,其中p,p′-DDT去除率较高,为49.7%;阴极土壤pH随电动试验时间的增加而增大,其最大值为11.63,阳极土壤表现相反,其最小值为-1.71;电流和土壤温度随电动试验时间的增加呈先增后减的趋势,其峰值均出现在第8小时.研究显示,不同DDTs异构体在电动力学作用下的迁移去除效果存在差异,并且电动过程会引起土壤理化性质改变.      

Bioaugmentation of DDT-contaminated soil by dissemination of the catabolic plasmid pDOD

A plasmid transfer-mediated bioaugmentation method for the enhancement of dichlorodiphenyltrichloroethane (DDT) degradation in soil was developed using the catabolic plasmid pDOD from Sphingobacterium sp. D-6. The pDOD plasmid could be transferred to soil bacteria, such as members of Cellulomonas, to form DDT degraders and thus accelerate DDT degradation. The transfer efficiency of pDOD was affected by the donor, temperature, moisture, and soil type. Approximately 50.7% of the DDT in the contaminated field was removed 210 days after the application of Escherichia coli TG I (pDOD-gfp). The results suggested that seeding pDOD into soil is an effective bioaugmentation method for enhancing the degradation of DDT.     

干法水泥回转窑共处置滴滴涕废物的示范工程

利用新型干法悬浮预热预分解回转窑对滴滴涕废物进行共处置研究.滴滴涕废物以1t/h的投加速率,从窑尾烟室加入窑系统,比较了未处置废物工况和处置滴滴涕废物工况下烟气、生料、熟料、飞灰中滴滴涕和二 浓度.结果表明:在投加滴滴涕废物工况下,滴滴涕得到有效分解,其销毁率和销毁去除率分别为99.99953%和99.999982%;在处置滴滴涕废物前后烟气二 浓度没有发生明显变化,其浓度排放远低于水泥窑废物处置要求低于0.1ng-ITEQ/Nm3的水平;该水泥生产工艺系统在处置滴滴涕废物时,二 烟气排放因子为0.0396μg/t熟料,熟料排放因子为0.220μg/t;滴滴涕废物处置对炉窑生产,运行工况参数和常规污染排放没有显著影响.