修复PAHs复合污染体系的高效菌群构建及降解特性

从污染环境分离和筛选得到了9株PAHs降解菌,以其为基本菌种,构建高效修复PAHs复合污染体系的菌群(D)。采用平板稀释涂布计数法对降解体系中菌群(D)的动态结构进行了解析,数据显示菌群内的微生物在降解过程中能相互协作,发挥稳定且高效的降解作用。实验进一步研究了菌群D对单一PAH和混合PAHs的降解特性,结果表明,无论对单一PAH还是混合PAHs,菌群D均具有较强的降解能力。当降解历时6 d,菌群D能使40 mg/L的单一PAH平均降解85.8%,使60 mg/L的混合PAHs平均去除89.4%。菌群D在多环芳烃复合污染体系的生物修复方面具有潜在的应用价值。     

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。     

机油高效降解菌群筛选及降解效果初探

从多处受石油污染的土壤中经过初步筛选、混合驯化得到以机油为唯一碳源进行生长代谢的混合菌群.利用此混合菌群进行的降解实验结果表明,该菌群对高浓度机油废水具有较强的降解能力,初始含机油约2.0 g/L的人工废水.接种量为0.1%(菌体湿重/培养液体积),经过7 d的降解,机油可降至403 mg/L,降解率达81.4%;对不同浓度机油废水的降解实验结果表明,在静态实验条件下,机油质量浓度在不高于1 000 mg/L(含1 075 mg/L),混合菌群在降解过程中能自行从降解产酸的不良环境中恢复,机油质量浓度在2 000 mg/L以上,初期产酸较多,pH下降幅度较大,在7 d的周期内,废水pH无法恢复,说明在降解后期仍有大量有机酸积累而未被彻底降解;与葡萄糖共基质的降解实验结果表明,经过7 d的降解.不超过150 mg/L,的葡萄糖与1 000 mg/L机油组成的共基质体系中,机油降解基本不受葡萄糖加入的影响,但可加强早期的降解速率.而葡萄糖高于150 mg/L时,则会对混合菌群的除油率产生抑制,抑制程度随着葡萄糖浓度的提高而加大.     

盐环境下降解菌群对芘的降解特性研究

以芘为多环芳烃(PAHs)的代表物,利用1.0%盐度的无机盐培养基从石油污染土壤中富集出高效嗜盐PAHs降解菌群。通过DNA测序鉴定,菌群中对芘起重要降解作用的是Rhodanobacter、Pseudomonas、Mycobacterium,3者碱基比例达到31.82%。14d内,萘、菲、荧蒽、芘、苯并[a]芘5种PAHs的挥发损耗均可忽略不计。筛选得到的菌群降解芘的最佳条件为:酵母粉质量浓度为120mg/L,盐度不超过1.0%,无需额外添加甲基-β-环糊精。筛选出的降解菌群对芘的最佳降解条件可用于降解萘、菲、荧蒽和苯并[a]芘等其他PAHs,但随着PAHs环数增加,分子量增大,降解率降低。在最佳条件下降解14d时,萘、菲、荧蒽、芘、苯并[a]芘5种PAHs的降解率可分别达100.00%、85.48%、51.92%、56.28%、50.45%。     

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。     

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。     

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。     

高效降氰菌群的构建及降解特性

从筛选到的降氰菌中构建出了优于单一菌种降氰活性的复配菌群CNR.研究了该复合菌群的生长条件,探讨了温度、pH、接种量、氰化物初始浓度及降解时间等因素对CNR降氰的影响.研究表明,复配菌群CNR适应碱性环境,可降解高浓度氰化物(CN-11 000 mg/L),并对金属氰化物和脂肪族腈具有极强的降解能力.在有氧、pH 11、33 ℃和接种量10%条件下.含CN-11 000 ng/L培养液经60 h降解后,CN-1浓度降为0.49 mg/L,降氰率高达99.96%,达到国家一级排放标准.     

耐受重金属的芘降解真菌的筛选及芘降解动力学研究

为了提高复合污染土壤修复的微生物资源的丰富度,为混合菌群修复污染土壤积累资料,利用多环芳烃-重金属双抗培养基在污染土壤中筛选得到一株对Cu和Cd有高耐受性的芘降解真菌,经分子生物学鉴定为米曲霉。探究了米曲霉对芘污染水体的降解效果及对重金属Cu和Cd的耐受程度,利用缺乏生长基质的毒性抑制动力学模型对芘单基质降解过程进行了拟合,以期为后续共代谢、固定化的研究及实际工程应用提供一定的理论支撑。结果表明:(1)米曲霉以芘为单基质代谢时,降解率为33%;(2)米曲霉对重金属Cu和Cd的耐受浓度分别为500 mg/L和50 mg/L,分别高出国家土壤重金属二级标准5倍和83倍;(3)米曲霉对单基质芘的降解符合Crridle提出的毒性抑制动力学简化模型Sc=Sc0·Tb*cX0(1-e-bt)(R2=0.9237)。芘初始浓度Sc0=80 mg/L,米曲霉投加量X0=85 mg/L时,数值拟合得到内源呼吸常数b=0.027,生物转化量Tb*c=0.2875。该米曲霉对单基质芘及重金属Cu和Cd表现出一定的降解性能及耐受性能,故可经过适当强化后作为多环芳烃-重金属污染土壤的微生物修复菌种。     

喹啉降解菌的降解效果考察

为了探索石油污染土壤中含氮杂环化合物的降解情况,在考察石油污染土壤理化性质的基础上,选择喹啉作为目标污染物,采用选择性富集培养的方法,从45份石油污染土壤样品中,分离得到155株降解喹啉污染物的高效降解菌株,从中选择降解效率较高的2株喹啉降解菌命名为Q5和Q24,进行喹啉的降解性能研究,比较了单一优势菌株、人工复合菌群和土壤中的自然菌群对喹啉的降解情况。实验结果表明,石油污染土壤中自然菌群对喹啉的降解效果好于单一的优势菌株和人工复合菌群。     

PAH dissipation in a contaminated river sediment under oxic and anoxic conditions

A batch experiment was conducted to compare PAH degradation in a polluted river sediment under aerobic and anaerobic conditions, and to investigate whether input of fresh organic material (cellulose) could enhance such degradation. All measurements were checked against abiotic control treatments to exclude artifacts of sample preparation and non-biological processes like aging. Three- and four-ring PAHs could be degraded by the indigenous microbial community under aerobic conditions, but anaerobic metabolism based on iron and sulphate reduction was not coupled with PAH degradation of even the simplest 3-ring compounds like phenanthrene. Cellulose addition stimulated both aerobic and anaerobic respiration, but had no effect on PAH dissipation. We conclude that natural attenuation of PAHs in polluted river sediments under anaerobic conditions is exceedingly slow. Dredging and biodegradation on land under aerobic conditions would be required to safely remediate and restore polluted sites.     

Remediation of PAH contaminated soils: application of a solid-liquid two-phase partitioning bioreactor

The feasibility of a two-step treatment process has been assessed at laboratory scale for the remediation of soil contaminated with a model mixture of polycyclic aromatic hydrocarbons (PAHs) (phenanthrene, pyrene, and fluoranthene). The initial step of the process involved contacting contaminated soil with thermoplastic, polymeric pellets (polyurethane). The ability of three different mobilizing agents (water, surfactant (Biosolve) and isopropyl alcohol) to enhance recovery of PAHs from soil was investigated and the results were compared to the recovery of PAHs from dry soil. The presence of isopropyl alcohol had the greatest impact on PAH recovery with approximately 80% of the original mass of PAHs in the soil being absorbed by the polymer pellets in 48 h. The second stage of the suggested treatment involved regeneration of the PAH loaded polymers via PAH biodegradation, which was carried out in a solid-liquid two-phase partitioning bioreactor. In addition to the PAH containing polymer pellets, the bioreactor contained a microbial consortium that was pre-selected for its ability to degrade the model PAHs and after a 14 d period approximately 78%, 62% and 36% of phenanthrene, pyrene, and fluoranthene, respectively, had been desorbed from the polymer and degraded. The rate of phenanthrene degradation was shown to be limited by mass transfer of phenanthrene from the polymer pellets. In case of pyrene and fluoranthene a combination of mass transfer and biodegradation rate might have been limiting.     

Application of magnetite-activated persulfate oxidation for the degradation of PAHs in contaminated soils

In this study, feasibility of magnetite-activated persulfate oxidation (AP) was evaluated for the degradation of polycyclic aromatic hydrocarbons (PAHs) in batch slurry system. Persulfate oxidation activated with soluble Fe(II) (FP) or without activation (SP) was also tested. Kinetic oxidation of PAHs was tracked in spiked sand and in aged PAH contaminated soils at circumneutral pH. Quartz sand was spiked with: (i) single model pollutant (fluorenone) and (ii) organic extract isolated from two PAH contaminated soils (H and NM sampled from ancient coking plants) and was subjected to oxidation. Oxidation was also performed on real H and NM soils with and without an extraction pretreatment. Results indicate that oxidation of fluorenone resulted in its complete degradation by AP while abatement was very low (<20%) by SP or FP. In soil extracts spiked on sand, significant degradation of 16 PAHs was observed by AP (70-80%) in 1 week as compared to only 15% by SP or FP systems. But no PAH abatement was observed in real soils whatever the treatment used (AP, FP or SP). Then soils were subjected to an extraction pretreatment but without isolation of organic extract from soil. Oxidation of this pretreated soil showed significant abatement of PAHs by AP. On the other hand, very low degradation was achieved by FP or SP. Selective degradation of PAHs was observed by AP with lower degradation efficiency towards high molecular weight PAHs. Analyses revealed that no by-products were formed during oxidation. The results of this study demonstrate that magnetite can activate persulfate at circumneutral pH for an effective degradation of PAHs in soils. However, availability of PAHs and soil matrix were found to be the most critical factors for degradation efficiency.     

Effects of soil organic matter on the development of the microbial polycyclic aromatic hydrocarbons (PAHs) degradation potentials

The microbial activity in soils was a critical factor governing the degradation of organic micro-pollutants. The present study was conducted to analyze the effects of soil organic matter on the development of degradation potentials for polycyclic aromatic hydrocarbons (PAHs). Most of the degradation kinetics for PAHs by the indigenous microorganisms developed in soils can be fitted with the Logistic growth models. The microbial activities were relatively lower in the soils with the lowest and highest organic matter content, which were likely due to the nutrition limit and PAH sequestration. The microbial activities developed in humic acid (HA) were much higher than those developed in humin, which was demonstrated to be able to sequester organic pollutants stronger. The results suggested that the nutrition support and sequestration were the two major mechanisms, that soil organic matter influenced the development of microbial PAHs degradation potentials.     

The lack of microbial degradation of polycyclic aromatic hydrocarbons from coal-rich soils

Analytical techniques used to assess the environmental risk of contamination from polycyclic aromatic hydrocarbons (PAHs) typically consider only abiotic sample parameters. Supercritical fluid extraction and sorption enthalpy experiments previously suggested slow desorption rates for PAH compounds in two coal-contaminated floodplain soils. In this study, the actual PAH availability for aerobic soil microorganisms was tested in two series of soil-slurry experiments. The experimental conditions supported microbial degradation of phenanthrene if it was weakly sorbed onto silica gel. Native coals and coal-derived particles in two soils effectively acted as very strong sorbents and prevented microbial PAH degradation. The long history of PAH exposure and degree of coal contamination apparently had no influence on the capability of the microbial soil community to overcome constraints of PAH availability. Within the context of the experimental conditions and the compounds chosen, our results confirm that coal-bound PAHs are not bioavailable and hence of low environmental concern.     

Fate of polycyclic aromatic hydrocarbons during composting of lagooning sewage sludge

The fate of 16 polycyclic aromatic hydrocarbons (PAHs), targeted by the USEPA agency, has been investigated during composting of lagooning sludge. Composting shows efficient decrease of the content and the bioavailability of each PAH. Biodegradation and sorption are suggested as the main mechanisms contributing to this decrease. During the stabilization phase of composting, extensive microbial degradation of PAHs, mainly those with a low number of aromatic rings, was achieved following development of intense thermophilic communities. However, partial sorption of PAH to non-accessible sites temporarily limits the mobility mainly of PAHs with a high number of aromatic rings plus acenaphthene and acenaphthylene, and allows them to escape microbial attack. During the maturation phase, the development of a mesophilic population could play an important role in the degradation of the remaining PAH. During this phase of composting, PAH sequestration and binding of their oxidative metabolites within new-formed humic substances might also explain PAH decrease at the end of composting. The tendency of change of content or bioavailability of various PAH compounds during composting is found to be strongly related to the number of their aromatic rings, their molecular weight and structure.     

PAH dissipation in spiked soil: impacts of bioavailability, microbial activity, and trees

While trees have demonstrated potential in phytoremediation of several organic contaminants, little is known regarding their ability to impact the common soil contaminant PAHs. Several species of native North American trees were planted in soil artificially contaminated with three PAHs. Plant biomass, PAH dissipation, and microbial mineralization were monitored over the course of one year and environmental conditions were allowed to follow typical seasonal patterns. PAH dissipation and mineralization were not affected by planting. Extensive and rapid loss of PAHs was observed and attributed to high bioavailability and microbial activity in all treatments. The rate of this loss may have masked any significant planting effects. Anthracene was found to be more recalcitrant than pyrene or phenanthrene. Parallel soil aging studies indicated that sequestration to soil components was minimal. Contrary to common inferences in literature, amendment with decaying fine roots inhibited PAH degradation by the soil microbial community. Seasonal variation in environmental factors and rhizosphere dynamics may have also reduced or negated the effect of planting and should be taken into account in future phytoremediation trials. The unique root traits of trees may pose a challenge to traditional thought regarding PAH dissipation in the rhizosphere of plants.     

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.     

Evidence for bioaccumulation of PAHs within internal shoot tissues by a halophytic plant artificially exposed to petroleum-polluted sediments

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental pollutants of natural and anthropic origins. Despite their poor water solubility, they can be taken up and bioaccumulated by plants. This study was aimed at determining whether the PAHs present in sediments artificially polluted by heavy fuel oil are transferred to shoots of a coastal and edible plant, Salicornia fragilis Ball et Tutin. Bioaccumulation was quantified after a one-week exposure to sediments polluted with 0.2%, 2% and 20% fuel oil (w/w) and over a six-week monitoring at 0.2%. Quantification by GC-MS of PAH amounts in plants and sediments evidenced a bioaccumulation in the shoots by a soil-to-plant transfer through the root system. This bioaccumulation depended on the duration of exposure and on the substratum contamination. PAHs distributions in plants and sediments looked alike with a predominance of low- and medium-weight hydrocarbons. Moreover, high-weight PAHs were also detected in the upper part of plants.     

Combined UV-biological degradation of PAHs

The UV-photolysis of PAHs was tested in silicone oil and tetradecane. In most cases, the degradation of a pollutant provided within a mixture was lower than when provided alone due to competitive effects. With the exception of anthracene, the larger pollutants (4- and 5-rings) were always degraded first, proving that UV-treatment preferentially acts on large PAHs and thereby provides a good complement to microbial degradation. UV-photolysis was also found to be suitable for treatment of soil extract from contaminated soils. The feasibility of UV-biological treatment was demonstrated for the removal of a mixture of phenanthrene and pyrene in silicone oil. UV-irradiation of the silicone oil led to 83% pyrene removal but no phenanthrene photodegradation. Subsequent treatment of the oil in a two-phases partitioning bioreactor (TPPB) system inoculated with Pseudomonas sp. was followed by complete phenanthrene biodegradation but no further pyrene removal. Totally, the combined process allowed 92% removal of the PAH mixture. Further work should focus on characterizing the photoproducts formed and studying the influence of the solvent on the photodegradation process.