污染土壤中苯并（a）芘的微生物降解

本文以B(a)P污染土壤为处理对象,进行了土壤微生物群体及单一菌对B（a)P的降解试验,结果表明,鲜土中的微生物降解力＞风干土的微生物降解力。土壤微生物群体比单一微生物降解B（a)P效果好。真菌的降解能力强于细菌。2209号镰刀菌降解B（a)P的速率最快,为高效降解菌。     

阿特拉津土壤污染修复菌剂载体材料的筛选与应用

阿特拉津是长残留除草剂,其环境行为和生物修复技术已成为有机污染控制领域的研究热点。以廉价的高岭土、凹凸棒土和腐殖酸为载体材料,采用正交实验,把功能菌存活率作为目标性状,参考材料成球率,筛选出性能较好的高岭土、凹凸棒土和腐殖酸质量配比3种,分别为1∶0.5∶0.5(A3B2C1)、0.5∶0∶0.5(A2B3C1)和1∶0∶1(A3B1C2);在温度和紫外线耐受力实验中,A3B2C1材料配比更能够有效提升功能菌在高温和紫外线作用下的存活率,即载体中高岭土、凹凸棒土和腐殖酸质量比为1∶0.5∶0.5时效果最佳;利用A3B2C1载体材料配比制备菌剂,进行室内土壤修复实验,35 d时0.1%和0.5%载体菌剂添加量修复土壤中阿特拉津完全降解,而2个游离菌修复土壤中残留率均16%,0.1%载体菌剂添加量修复过程中土壤微生物Shannon多样性指数和均匀度变化幅度较其他修复方式小,有利于土壤微生物生态系统的平衡,因此0.1%载体菌剂添加量修复效果为最优。     

多环芳烃降解菌的筛选及其在焦化场地污染土壤修复中的应用

经过富集、分离和纯化,从沈阳某焦化厂多环芳烃(PAHs)污染土壤中获得7株菌株B1~B7。通过初步降解实验和血平板实验,发现B4、B5、B7在15d时对PAHs总降解率均高于40%,为高效PAHs降解菌,B2为高效表面活性剂产生菌。将B4、B5、B7分别与B2等质量混合后对PAHs进行降解,发现添加B2可提高PAHs总降解率,B4+B2对PAHs的总降解率最大,在9d时平均值达到45.9%。经形态观察和16SrRNA基因序列比对,鉴定B2和B4分别归为假单胞菌属(Pseudomonas sp.)和芽孢杆菌属(Bacillus sp.)。接种B4+B2进行微生物修复实验,结果表明,接种B4+B2对PAHs污染土壤的微生物修复有明显的强化作用,在60d时PAHs总降解率达到48.1%;接种B4+B2对中环(4、5环)PAHs降解率的提高尤为明显,7种中环PAHs的平均降解率比不接种菌株的对照组提高29.6百分点。     

氧化石墨烯强化铜绿假单胞菌降解污染土壤多环芳烃的效果及其机制研究

微生物修复技术具有经济绿色、环境友好等特征,已成为多环芳烃(PAHs)污染土壤的主要修复手段之一。然而,针对经历长期老化的污染场地土壤,微生物修复效率偏低,生物强化技术亟待进一步提高。以PAHs高效降解菌铜绿假单胞菌(Pseudomonas aeruginosa,PAE)为对象,研究了新型碳质纳米材料氧化石墨烯(GO)对PAE生长和PAHs降解的影响,探讨了GO强化PAE降解土壤PAHs的效果及其机制。结果显示：(1)50～100 mg/L GO可以显著促进PAE的生长和胞外聚合物(EPS)的分泌。(2)PAE及GO(100 mg/kg)的添加显著促进了老化土壤中PAHs的降解。(3)GO添加前期,土著微生物群落丰度下降,PAE丰度显著增加;处理后期,土壤细菌群落丰度恢复至对照组水平。适宜浓度GO的添加可以影响土壤微生物的多样性和丰度,促进PAHs的降解,然而,修复后期GO的影响力下降,土壤微生物群落呈现出“扰动—恢复”模式。研究结果有助于深入理解GO对环境微生物的效应,为PAHs污染土壤的微生物修复提供新思路。     

电子受体对深层土壤中石油烃缺氧降解的影响

微生物降解是处理土壤中石油烃 (PHC)污染的有效技术,目前对PHC微生物降解的研究多集中在好氧条件下,对PHC缺氧微生物降解的研究较少,PHC缺氧降解规律尚不清楚。以PHC污染的深层土壤为对象,探究不同质量分数 (500、1 500、5 000 mg·kg⁻¹)的硫酸盐、硝酸盐或混合电子受体对土壤中土著微生物丰度、群落结构以及PHC缺氧降解的影响规律。结果表明,150 d缺氧培养后,添加相同种类电子受体的土壤处理中细菌丰度、潜在PHC降解菌 (变形菌门和厚壁菌门)丰度随电子受体的质量分数增加而增加;添加相同质量分数的不同种类电子受体土壤处理中细菌丰度、潜在PHC降解菌丰度从高到低分别为硝酸盐、混合电子受体、硫酸盐。添加相同种类电子受体的土壤处理中ΣPHC (C₁₀~C₃₀)和C1 (C₁₀~C₁₆)、C2 (C₁₇~C₂₃)、C3 (C₂₄~C₃₀)组分的降解率随着加入电子受体质量分数增加而增加;相同质量分数的不同种类电子受体土壤处理中ΣPHC和C1、C2、C3组分的降解率从高到低分别为硝酸盐、混合电子受体、硫酸盐。土壤中PHC缺氧降解率与细菌丰度、潜在PHC降解菌丰度均存在正相关关系。研究结果可为石油烃污染土壤的修复技术研发提供技术支持。     

化学预氧化-生物强化-生物刺激对土壤中菲降解的联合效应

通过室内实验,探究了低浓度过硫酸盐预氧化耦合生物强化或生物刺激技术处理下土壤中菲的降解率和修复效应。结果表明,浓度为0.1 mmol·g~(-1)、温度为50℃热活化的过硫酸钠对土壤中菲7 d的降解率为22.7%。预氧化后,加入高效降解菌和营养物质,强化微生物对菲的降解,继续培育21 d,最终降解率较第7天可提高8.08%～18.59%。同时添加高效降解菌和营养物质N,对土壤中菲的降解促进作用最强,最终降解率可达41.29%,较仅进行化学氧化的对照组和仅进行微生物降解的对照组分别提高17.44%和22.86%,较预氧化后不进行微生物强化的对照组提高12.9%。降解期间,土壤微生物数量和pH呈先下降,后上升趋势,最终维持在相对稳定水平。相关性分析结果表明,土壤中菲的降解率与氧化剂和营养物质N的添加呈显著正相关,土壤微生物数量与pH呈正相关,与氧化剂呈负相关,土壤pH与氧化剂及营养物质P呈负相关。研究结果证实了化学预氧化耦合生物强化和生物刺激技术能有效促进微生物对菲污染土壤的修复。     

黑土微生物群落对副球菌属(Paracoccus sp.)QD15-1修复邻苯二甲酸二甲酯污染土壤的响应

采用Illumina MiSeq高通量测序法探究黑土微生物群落对副球菌属(Paracoccus sp.)QD15-1修复邻苯二甲酸二甲酯(DMP)污染土壤的响应。结果表明,副球菌属QD15-1在7d内可快速有效降解土壤中的DMP,平均降解率为94.7%,并能保持稳定;Shannon、Sobs指数显示,土壤中微生物多样性、丰富性受到DMP的影响;土壤中的优势门为放线菌门(Actinobacteria),占总菌门数的50%以上;鞘氨醇单胞菌属(Sphingomonas)是第一优势菌属,第二优势菌属为类诺卡氏菌属(Nocardioides)。     

一株多环芳烃降解菌的筛选及其降解特性

微生物修复是治理土壤多环芳烃(polycyclic aromatic hydrocarbons, PAHs)污染的主要方法,而高效降解菌筛选是微生物修复技术的重要基础。从北京焦化厂土壤中筛选分离得到一株PAHs降解菌Q3,通过生理生化和16S rDNA等分析手段鉴定其为Rhodococcus rhodochrous。结果表明：该菌株对芘的耐受能力较强,可降解初始浓度为200 mg·L⁻¹的芘;该菌株具有降解广谱性,可利用苯并[a]芘、苯并[b]荧蒽、二苯并[a,h]蒽、苯并[g,h,i]苝等9种PAHs为唯一碳源进行代谢,特别是对苯并[a]芘等高环PAHs具有较好的降解效果;此外,该菌株可有效降解模拟液中的混合PAHs,并且对野外被PAHs长期污染的土壤具有较好的强化修复效果。投加菌株处理后的处理组与对照组相比,土壤PAHs总去除率提高了24%。以上结果表明该菌株对环境中被PAHs污染的土壤具有较好的强化修复潜力,可为PAHs污染土壤的微生物修复技术提供技术参考。     

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。     

规模化奶牛养殖粪水还田对土壤理化性质及微生物群落特征的影响

规模化奶牛养殖粪水还田可实现资源化利用,但其对土壤环境的影响尚待明晰。通过完全随机区组设计试验研究了粪水还田量分别为0、0.32、0.64、0.96 t/m²时对土壤理化性质及微生物群落特征的影响。结果表明：养殖粪水还田可以提高土壤养分和作物生物量,最合适的粪水还田量应为0.64 t/m²,此时对土壤微生物的丰富度和多样性影响也最小。土壤细菌中的优势细菌属斯克尔曼氏菌属(Skermanella)可生物固氮,减少土壤氮素流失;芽孢杆菌属(Bacillus)能分泌纤维素酶,有利于土壤中纤维素类的降解。子囊菌门(Ascomycota)为最主要的优势真菌门,它们有助于植物纤维素和木质素的降解。     

利用生物炭负载微生物修复石油烃-镉复合污染土壤

自四川省长宁某石油烃-重金属镉复合污染土壤中,筛选出1株具有重金属镉(Cd)抗性和石油烃降解能力的菌株,采用吸附法将菌株固定于玉米芯生物炭上制备成固定化微生物,并探讨固定化微生物技术(IMT)对土壤石油烃的降解效率以及重金属固定化效果。鉴定结果表明,成功筛选出的具有Cd抗性的石油烃降解菌经鉴定属于柠檬酸杆菌属(Citrobacter sp.)。实验结果表明：固定化微生物修复石油烃-重金属镉复合污染土壤60 d后,固定化微生物对石油烃降解率达51.25%,显著高于游离菌组(40.44%)、生物炭组(31.11%)和空白组(15.18%)(P<0.05);同时,固定化微生物能够高效固定土壤重金属Cd,使其从可交换态、有机结合态向残渣态转变,可使其残渣态与初始土壤相比增加6.68倍;固定化微生物组的土壤pH较初始出现轻微下降,但维持于8.42~8.75;此外,固定化微生物可显著增加土壤细菌数量(2.48×10⁸ cfu·g⁻¹)(P<0.05),土壤脱氢酶活性、过氧化氢酶活性以及多酚氧化酶活性(P<0.05)。因此,采用生物炭固定化微生物技术,可在高效降解石油烃的同时,提高重金属Cd固定化效果。该研究结果可为IMT技术修复石油烃-镉复合污染土壤提供参考。     

土著反硝化菌对巢湖流域稻田泥土脱氮效果的影响

生物修复技术削减途经土壤中的硝态氮是一种环境友好且生态效益高的方法。构建了含不同填料的A (土壤) 、B (土壤+木屑) 、C (土壤+木屑+蜡样芽孢杆菌) 、D (土壤+木屑+土著反硝化菌) 4种渗透反应柱,将从土壤填料中筛选出单株具有良好反硝化性能的土著反硝化菌 (Pseudomonas sp. P10) 和来源于湖泊底泥的反硝化菌蜡样芽孢杆菌分别接种至渗透反应柱,探究土著反硝化菌对巢湖流域同源水稻田土壤填充渗透反应柱脱氮效果的影响。结果表明,向反应柱中接种土著反硝化菌Pseudomonas sp. P10可缩短反应柱去除NO₃⁻-N的启动时间,填料为土壤与木屑的B柱对NO₃⁻-N去除率可达96.05%。 接种土著反硝化菌的D柱出现了NH₄⁺-N积累现象, B、C、D柱中积累的[NO₂⁻-N]均低于2.4 mg·L⁻¹。在30 mg·L⁻¹和50 mg·L⁻¹的NO₃⁻-N进水条件下,各反应柱对TN去除率为50%~70%。各反应柱出水TOC符合水质标准。接种土著反硝化菌的D柱中优势菌种为Pseudomonas属细菌,C柱中接种的蜡样芽孢杆菌并没能成功地与原有微生物群落竞争。接种土著反硝化菌对土壤反硝化能力有一定强化作用,但随着时间的延长,在充足可利用碳源条件下,没有接种微生物的反应柱更具有反硝化脱氮优势。该研究结果可为提升土壤对外源氮素的削减和湖泊氮素污染的控制提供参考,也为湖库淤积物的资源化利用及治污情况下生态系统的稳态维持和可持续的系统动力学提供依据。     

Effect of vermicomposting on calcium, sulphur and some heavy metal content of different biodegradable organic wastes under liming and microbial inoculation

A study was conducted to evaluate the changes in total calcium and sulphur and some heavy metal (Zn, Cu, and Pb) concentration of different organic wastes affected by liming and microorganism inoculation. Vermicomposting was an effective technology for disposal of organic substrates like municipal solid wastes (MSW), possessing comparatively higher concentration of heavy metals. The addition of lime in initial organic substrates significantly (P ≤ 0.05) increased total calcium and total sulphur content of vermicomposts. Inoculation of microorganisms significantly (P ≤ 0.05) reduced the heavy metal content of final products as compared to control. Fungal strains were comparatively more effective in detoxification of heavy metals than B. polymyxa.     

Alginate immobilized enrichment culture for atrazine degradation in soil and water system

An atrazine degrading enrichment culture, a consortium of bacteria of genus Bacillus along with Pseudomonas and Burkholderia, was immobilized in sodium alginate and was used to study atrazine degradation in mineral salts medium (MSM), soil and wastewater effluent. Sodium alginate immobilized consortium, when stored at room temperature (24 ± 5°C), was effective in degrading atrazine in MSM up to 90 days of storage. The survival of bacteria in alginate beads, based on colony formation unit (CFU) counts, suggested survival up to 90 days and population counts decreased to 1/5^th on 120 days. Comparison of atrazine degrading ability of the freely suspended enrichment culture and immobilized culture suggested that the immobilized culture took longer time for complete degradation of atrazine as a lag phase of 2 days was observed in the MSM inoculated with alginate immobilized culture. The free cells resulted in complete degradation of atrazine within 6 days, while immobilized cells took 10 days for 100% atrazine degradation. Further, immobilized cultures were able to degrade atrazine in soil and wastewater effluent. Alginate beads were stable and effective in degrading atrazine till 3rd transfer and disintegrated thereafter. The study suggested that immobilized enrichment culture, due to its better storage and application, can be used to degrade atrazine in soil water system.     

Anaerobic degradation of nonylphenol in soil

This study investigated the effects of various factors on the anaerobic degradation of nonylphenol (NP) in soil. The results show that the optimal pH for NP degradation was 7.0 and that the degradation rate was enhanced when the temperature was increased. The addition of compost enhanced NP degradation. The individual addition of the electron donors lactate, acetate, and pyruvate inhibited NP degradation. The high-to-low order of NP degradation rates under three anaerobic conditions was sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The results show that sulfate-reducing bacteria, methanogen, and eubacteria are involved in the anaerobic degradation of NP, with sulfate-reducing bacteria being a major component of the soil. Of the anaerobic strains isolated from the soil samples, strain AT3 expressed the best ability to biodegrade NP.     

Anaerobic degradation of nonylphenol in soil

This study investigated the effects of various factors on the anaerobic degradation of nonylphenol (NP) in soil. The results show that the optimal pH for NP degradation was 7.0 and that the degradation rate was enhanced when the temperature was increased. The addition of compost enhanced NP degradation. The individual addition of the electron donors lactate, acetate, and pyruvate inhibited NP degradation. The high-to-low order of NP degradation rates under three anaerobic conditions was sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The results show that sulfate-reducing bacteria, methanogen, and eubacteria are involved in the anaerobic degradation of NP, with sulfate-reducing bacteria being a major component of the soil. Of the anaerobic strains isolated from the soil samples, strain AT3 expressed the best ability to biodegrade NP.     

Degradation of atrazine by an acclimatized soil fungal isolate

A fungal strain able to use atrazine (2-chloro-4-ethylamino-5-isopropylamino-1,3,5-triazine) as a source of nitrogen was isolated from a corn field soil that has been previously treated with the herbicide. This strain was purified and acclimatized to atrazine at a higher level in the laboratory. A supplemented N was required to trigger the reaction. Atrazine was degraded at a faster rate in inoculated mineral salt medium (MSM) than non-inoculated MSM. Within 20 days, nearly 34% of the atrazine was degraded in inoculated medium while only 2% of the herbicide was degraded in non-inoculated medium. Degradation of atrazine by the isolated fungal strain was also studied in sterile and non-sterile soil to determine the compatibility of the isolated strain with native microorganisms in soil. The degradation of atrazine was found to be more in inoculated sterile soil than in inoculated non-sterile soil. Cell free extract (CFE) of fungal mycelium degraded about 50% of the atrazine in buffer in 96 hours compared to the control. Four atrazine metabolites were isolated and characterized by LCMS. On the basis of morphological parameters the isolate was identified as Penicillium species. Results indicated that the microorganism may be useful for remediation of atrazine-contaminated soil.     

A comparative study of cadmium phytoextraction by accumulator and weed species

Phytoextraction has shown great potential as an alternative technique for the remediation of metal contaminated soils. The objective of this study was to investigate cadmium (Cd) phytoextraction ability of high biomass producing weeds in comparison to indicator plant species. The pot study conducted with 10 to 200 mg Cd kg(-1) soil indicated that Ipomoea carnea was more effective in removing Cd from soil than Brassica juncea. Among the five species, B. juncea accumulated maximum Cd, but I. carnea followed by Dhatura innoxia and Phragmytes karka were the most suitable species for phytoextraction of cadmium from soil, if the whole plant or above ground biomass is harvested. In the relatively short time of this experiment, I. carnea produced more than 5 times more biomass in comparison to B. juncea. There were significant differences (p < 0.05) between the shoot length and shoot mass of control and treated plants.     

The use of NTA and EDDS for enhanced phytoextraction of metals from a multiply contaminated soil by Brassica carinata

The potential of nine different species to grow in the presence of metals (As, Cd, Cu, Pb and Zn) and to accumulate them in the shoots was assessed for each metal separately by germination and root length tests, and successively by hydroponic experiments. Of the nine species tested, Brassica carinata was the species that accumulated the highest amounts of metals in shoots without suffering a significant biomass reduction. To further evaluate the potential of B. carinata for chelant-enhanced phytoextraction of a natural, multiply metal-polluted soil (As, Cd, Cu, Pb and Zn), both hydroponic and pot experiments were carried out with nitrilotriacetic acid (NTA) or (S,S)-ethylenediamine disuccinic acid (EDDS) as complexing agents. The hydroponic study with solutions containing the five metals together showed that accumulation of Cd, Cu, Pb and Zn in shoots was higher following EDDS addition compared to NTA. EDDS was more effective than NTA in desorbing Cu, Pb and Zn from the soil, whereas As and Cd were poorly extracted. B. carinata plants were grown for 4 weeks in the multiply metal-contaminated soil and then the soil was amended with 5 mmol kg(-1) NTA or EDDS. All plants were harvested 1 week after amendment. In comparison to NTA, EDDS was more effective in enhancing the concentrations of Cu, Pb and Zn in B. carinata shoots (2- to 4-fold increase compared to the control). One week after chelant addition, the DTPA-extractable metal concentrations in the polluted soil were lower in the EDDS treatment in comparison with the NTA amendment. Even though B. carinata showed a reduced growth and a relatively low metal uptake, it demonstrated the ability to survive and tolerate the presence of more metals simultaneously.     

Bioremediation of polycyclic aromatic hydrocarbon-contaminated saline-alkaline soils of the former Lake Texcoco

Polycyclic aromatic hydrocarbons (PAHs) such as phenanthrene, anthracene and Benzo[a]pyrene (BaP) are toxic for the environment. Removing these components from soil is difficult as they are resistant to degradation and more so in soils with high pH and large salt concentrations as in soil of the former lake Texcoco, but stimulating soil micro-organisms growth by adding nutrients might accelerate soil restoration. Soil of Texcoco and an agricultural Acolman soil, which served as a control, were spiked with phenanthrene, anthracene and BaP, added with or without biosolid or inorganic fertilizer (N, P), and dynamics of PAHs, N and P were monitored in a 112-day incubation. Concentrations of phenanthrene did not change significantly in sterilized Acolman soil, but decreased 2-times in unsterilized soil and >25-times in soil amended with biosolid and NP. The concentration of phenanthrene in unsterilized soil of Texcoco was 1.3-times lower compared to the sterilized soil, 1.7-times in soil amended with NP and 2.9-times in soil amended with biosolid. In unsterilized Acolman soil, degradation of BaP was faster in soil amended with biosolid than in unamended soil and soil amended with NP. In unsterilized soil of Texcoco, degradation of BaP was similar in soil amended with biosolid and NP but faster than in the unamended soil. It was found that application of biosolid and NP increased degradation of phenanthrene, anthracene and BaP, but to a different degree in alkaline-saline soil of Texcoco compared to an agricultural Acolman soil.