产漆酶真菌筛选及其对PAHs污染土壤修复的初步研究

真菌漆酶可以高效转化多环芳烃(PAHs),因此,产漆酶真菌在PAHs污染土壤修复中极具应用前景。根据漆酶可将愈创木酚氧化为红色物质的特性,成功从土壤中筛选出一株能够分泌漆酶的真菌菌株F-1,初步鉴定该菌为疣孢漆斑菌(Myrothecium verrucaria)。通过Plackett-Burman试验对菌株F-1的产酶能力进行了分析,发现特定培养条件组合可将其酶活提高近300倍,达5628 U L-1,表明F-1的漆酶活性受到环境条件的显著影响。应用菌株F-1对PAHs污染土壤进行了初步修复研究,结果表明,接种F-1对菲、荧蒽、芘、苯并(a)蒽、屈、苯并(b)荧蒽、苯并(k)荧蒽、苯并(a)芘、二苯并(a,h)蒽、苯并(g,h,i)苝、茚苯(1,2,3-cd)芘等11种PAHs均有不同程度的降解,提示产漆酶真菌在PAHs污染土壤修复中的应用潜力。     

东莞市土壤中多环芳烃的含量、代表物及其来源

2002年10~12月采集东莞市各地土壤样品64个,采用气相色谱-质谱仪对土壤中的16种优先控制的多环芳烃(PAHs)进行分析。结果显示,东莞市土壤中16种PAHs平均含量413μg/kg,含量较高的几种组分分别是菲、萤蒽、屈、苯并[b]萤蒽、芘。与PAHs总含量相关性较好的7种组分分别为屈、苯并[a]蒽、苯并[b]萤蒽、苯并[k]萤蒽、苯并[a]芘、茚并[1,2,3]芘、二苯并[a,h]蒽;所有相关系数R2均超过0.9000,可作为东莞市土壤中PAHs的代表物。北部平原水乡的PAHs污染明显高于南部丘陵山区、水乡上游保护区以及中部过渡带。化石燃料高温燃烧产物的大气沉降为东莞市土壤中PAHs的主要来源。东莞市农业土壤中PAHs含量显著低于天津市受污染严重的农业土壤。     

南京有机污染风险区农田土壤多环芳烃污染状况研究

采集南京地区不同有机污染风险区农田表层土壤,用超快速液相色谱仪检测样品中15种EPA优控的多环芳烃(PAHs)含量。结果表明,被检农田土壤多环芳烃总量分布于306.0~1251.3μg kg~(-1)之间,均值682.0μg kg~(-1),四环以上高环多环芳烃占较大比例(80%)。根据欧洲土壤质量标准,所检土壤样本已达污染水平。不同风险污染区农田土壤PAHs的含量由高至低为:钢铁工业区、有机垃圾处理区、化工工业区及炼油工业区。钢铁工业区附近主要的污染物为荧蒽、芘、屈和苯并[a]蒽,分别占到污染物总量的16%、13%、10%和10%。采用荧蒽/(荧蒽+芘)与茚并[1,2,3-cd]芘/(茚并[1,2,3-cd]芘+苯并[g,h,i]苝)比值对各地污染物来源进行分析,结果发现调查区域的PAHs污染物以燃烧源为主,生物质燃料为主要污染物,部分地区同时有石油燃烧污染。     

某氮肥厂场地土壤PAHs污染特征研究

采用现场采样及室内测试方法对广州某氮肥厂原料车间和油库区土壤中16种优控多环芳烃（PAHs）的含量进行调查研究,分析了EPAHs含量及其组成特征和垂直分布特征,并在此基础上进行了源解析。结果表明,分析样品中∑PAHs范围在10-7795μg·kg,原料车间土壤中的∑PAHs小于油库区土壤中的,菲、芘、荧蒽、并（b）荧蒽、苯并（a）芘为主要污染物;油库土壤0-40cm的样品中16种PAHs均有检出,∑PAHs和单体分布基本一致;原料车间土壤∑PAHs和单体浓度随着地面深度的增加而减少。通过对单组分比值（菲／蒽,荧蒽／芘）的分析可以看出油库区土壤中PAHs来源于石油和燃烧源,而原料车间污染源主要为燃烧源。     

多环芳烃高效降解菌的筛选

以多环芳烃(PAHs)菲、蒽、芘、■和苯并(a)芘为供试物,对土著混合菌和引进菌同时进行筛选实验。结果表明,引进菌和土著混合菌经过驯化后对菲、蒽、芘、■和苯并(a)芘均具有一定的降解能力。其中,在pH=6时,混合菌U03在48h内对5种PAHs的降解率均相对较高,分别为:72.38%;64.46%;65.77%;66.49%和64.77%,并且其的降解速率在各菌剂中同样最快,通过SPSS数理统计分析软件对数据进行处理后得出,混合菌U03可在较短时间内达到较好的降解目的。室内模拟试验证明混合菌U03具有较强的降解PAHs的能力,混合菌的协同作用有利于污染土壤中PAHs的降解。     

小麦秸秆生物炭对石油烃污染土壤的修复作用

以小麦秸秆为原材料,在300℃下缺氧裂解3、6、8 h制备生物炭,比较了3种生物炭的产率、pH值、灰分以及C、H、N元素含量,表征了300℃、6 h生物炭的表面形态,并用其作为修复材料,对大港油田的石油污染土壤进行修复。结果表明,随裂解时间的延长,生物炭产率下降,pH值升高,灰分含量增加,H/C值下降,但产率、pH值、灰分和H/C值都是从3 h到6 h差异显著,6 h到8 h差异不显著。C元素含量先升高后下降。石油污染土壤经生物炭修复14 d和28d后,总石油烃降解率分别为45.48%和46.88%,均显著高于对照组。修复14 d后土壤中的萘、苊、苯并[a]蒽、屈、苯并[b]荧蒽、苯并[k]荧蒽、苯并[a]芘、茚并[1,2,3-cd]芘也都有不同程度的下降,其中苯并[a]芘含量下降幅度达98.18%,其他几种PAH的降解率也都高于对照组,28 d后这些PAH的含量又有上升趋势。这说明小麦秸秆裂解时间对生物炭的性质有影响;300℃、6h生物炭可以用来修复石油污染的土壤。     

低浓度环糊精强化Fenton试剂降解液相中茚并（1,2,3-cd）芘的试验研究

为了研究Fenton试剂对液相中茚并（1,2,3-cd）芘的降解效果,采用实验室内试验方法,考察并确定了H2O2浓度、FeSO4浓度、反应时间、pH值以及低浓度环糊精等因素对降解的影响。结果表明,使用Fenton试剂可以有效降解液相中的茚并（1,2,3-cd）芘。在降解过程中,对初始浓度为2.5mg·L-1的茚并（1,2,3-cd）芘,当H2O2浓度为20.0mmol·L-1,FeSO4浓度为3.0mmol·L-1时降解较为有效,去除率可达到62.88%,浓度过高或者过低都不利于反应的发生。Fenton氧化降解茚并（1,2,3-cd）芘的反应在60min内可以完成,并且pH值控制在3时更加有利于降解。反应体系中加入低浓度环糊精可以增加茚并（1,2,3-cd）芘在水相中的溶解度,当加入HPCD的浓度为3mg·L-1时,Fenton试剂对茚并（1,2,3-cd）芘的氧化去除率可由62.88%提高至71.24%。     

长江三角洲典型地区农田土壤多环芳烃分布特征与源解析

采集了长江三角洲地区苏州市的96个和嘉兴市的324个农田土壤样品,较为系统地分析了土壤中15种优控多环芳烃组成及含量,并运用比值法和主成分分析法判断了土壤中多环芳烃的来源。结果显示,苏州和嘉兴农田土壤中15种多环芳烃的总量范围分别为45.4~3 703μg kg-1和9.0~2 421μg kg-1,其平均值分别为312.5μg kg-1和152.4μg kg-1。苏州农田土壤苯并[a]芘含量平均值达21.4μg kg-1,超过了前苏联制定的土壤苯并[a]芘最高允许值(20.0μg kg-1)。苏州农田土壤中多环芳烃的含量从南部的吴江到北部的相城呈不断增加的趋势。嘉兴农田土壤多环芳烃含量远低于苏州,而在嘉兴毗邻吴江和上海的嘉善县土壤中多环芳烃含量较高。菲、荧蒽、苯并[b]荧蒽、芘、茚并[1,2,3-cd]芘是苏州土壤中多环芳烃的主要成分,而嘉兴土壤中多环芳烃的主要成分为菲、荧蒽、苯并[b]荧蒽、芘、屈。两个地区农田土壤多环芳烃的来源较为相似,主要为生物质和煤的燃烧以及石油源。     

蚯蚓和丛枝菌根真菌对南瓜修复多环芳烃污染土壤的影响

在温室盆栽实验条件下,研究接种AM(arbuscular mycorrhiza)真菌、蚯蚓(Eisenia fetida)对南瓜(Cucurbita moschata)修复3环以上多环芳烃(PAHs)污染农田土壤的影响,试验设置单接AM真菌、单接蚯蚓、双接AM真菌和蚯蚓、不接种的对照共4个处理,播种10周后收获。结果表明,接种AM真菌和蚯蚓促进AM真菌侵染南瓜,增加南瓜生物量;显著提高南瓜修复土壤中Phe(菲)、An(t蒽)、Py(r芘)、BkF(苯并(k)荧蒽)、BaP(苯并(a)芘)、BPe(r苯并(g,h,i)苝)等PAHs污染物的效率,促进南瓜高效地吸收3~5环PAHs,尤其是AM真菌和蚯蚓共同接种条件下对南瓜修复土壤效果最优;AM真菌利于南瓜转移根系吸收的高浓度PAHs化合物至地上部,降低PAHs对根系的胁迫,增强南瓜在高浓度PAHs污染土壤中存活,有利于南瓜应用于高浓度PAHs污染土壤的高效修复;蚯蚓对南瓜地下部吸持3~5环高分子量的PAHs化合物有积极作用。因此,选用的AM真菌和蚯蚓在土壤中具有协同作用,促进南瓜高效修复PAHs污染土壤。     

农药企业搬迁土地多环芳烃的风险评价

以常州市某农药厂搬迁土地为研究对象,在监测分析土壤中16种多环芳烃（PAHs）的基础上,对该区域土壤进行健康风险和生态风险评价。结果表明,研究区域土壤中∑PAHs的含量范围为0～1.546mg·kg-1,优势化合物中萘、菲等低环化合物含量大于高环的荧蒽、苯并[k]荧蒽和芘等化合物,且土壤中PAHs可能来源于石油源。健康风险评价结果在可接受的10-6～10-4范围内,而生态风险评价表明,尽管研究区域土壤中的多环芳烃不存在严重的生态风险,但是化合物苊和芴含量超出了风险评价低值（ER-L和ISQV-L）,存在着对生物的潜在危害。     

Formation of polycyclic aromatic hydrocarbons in northern and middle taiga soils

An integrated study of the qualitative and quantitative composition of polycyclic aromatic hydrocarbons (PAHs) in the atmospheric precipitation-soil-lysimetric water system was performed using high performance liquid chromatography. It was shown that the accumulation of low-molecular PAHs (phenanthrene, anthracene, fluoranthene, pyrene, benz[a]anthracene, and chrysene) in soils is due to the transformation of organic matter and the regional transport and deposition of PAHs with atmospheric precipitation on the underlying surface. High-molecular polyarenes (benz[b]fluoranthene, benz[k]fluoranthene, benz[a]pyrene, dibenz[a,h]anthracene, benzo[ghi]perylene, and indeno[1,2,3-cd]pyrene) mainly result from the decomposition of soil organic matter.     

Rapid, semimicro method for determination of polycyclic aromatic hydrocarbons in shellfish by automated gel permeation/liquid chromatography

A simple, rapid, easily automated method is described for the determination of polycyclic aromatic hydrocarbons (PAHs) in shellfish such as American lobster (Homarus americanus) and blue mussel (Mytilus edulis). PAHs are extracted from small amounts (1-8 g) of tissue by saponification in 1N ethanolic potassium hydroxide followed by partitioning into 2,2,4-trimethylpentane. This solution is evaporated just to dryness by rotary evaporation and the residue is dissolved in cyclohexane-dichloromethane (1 + 1) for gel permeation chromatography (GPC) on Bio-Beads SX-3. The GPC procedure is ideal as a screening method in the range 25-18 000 ng PAHs/g tissue. If individual PAH measurements are required, the appropriate GPC fraction is collected and PAHs are separated by reverse phase liquid chromatography (LC) with fluorometric detection. Individual PAHs at concentrations as low as 0.25-10 ng/g can be determined. Recoveries of added fluoranthene, pyrene, benz[a]anthracene, chrysene, benzo[e]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenz[a,h]anthracene, benzo[ghi]perylene, and indeno[1,2,3-cd]pyrene were quantitative, with relative standard deviations ranging from 0.0 to 16.9%.     

湿热灭菌和氯化汞灭菌对双液相体系中PAHs降解的影响

A two-liquid-phase(TLP) soil slurry system was employed to quantify the efficiencies of autoclaving and mercuric chloride sterilization in the dissipation of polycyclic aromatic hydrocarbons(PAHs).The fates of 11 PAHs(naphthalene,fluorene,phenanthrene,anthracene,fluoranthene,pyrene,benzo(a)anthracene,benzo(a)pyrene,benzo(b)fluoranthene,benzo(k)fluoranthene,dibenzo(a,h)anthracene) were recorded over 113 days of incubation.No microorganisms were detected in the HgCl 2-sterilized soil slurries during the whole incubation period,indicating very effective sterilization.However,about 2%-36% losses of PAHs were observed in the HgCl 2 sterilized slurry.In contrast to the HgCl 2-sterilized soil slurry,some microorganisms survived in the autoclaved soil slurries.Moreover,significant biodegradation of 6 PAHs(naphthalene,fluorene,phenanthrene,anthracene,fluoranthene and pyrene) was observed in the autoclaved soil slurries.This indicated that biodegradation results of PAHs in the soil slurries,calculated on basis of the autoclaved control,would be underestimated.It could be concluded that the sterilization efficiency and effectiveness of HgCl 2 on soil slurry was much higher than those of autoclaving at 121℃ for 45 min.     

Polycyclic aromatic hydrocarbon profile analysis of high-protein foods, oils, and fats by gas chromatography.

A method is described for the determination of polycyclic aromatic hydrocarbons (PAHs) with 3-7 rings in (I) meat, poultry, fish, and yeast; and (II) oils and fats. The extraction of PAHs from group I is incomplete, and, therefore, group I samples must be dissolved homogeneously by saponification in 2N methanolic potassium hydroxide. The PAHs are concentrated by liquid-liquid extraction (methanol-water-cyclohexane, N,N - dimethylformamide - water-cyclohexane) and by column chromatography on Sephadex LH 20. The PAHs are separated by high-performance gas-liquid chromatography (GLC) with columns containing 5% OV-101 on Gas-Chrom Q and estimated by integration of the flame ionization detector signals in relation to an internal standard (3,6-dimethylphenanthrene and/or benzo(b)chrysene). The sensitivity is significantly higher than that obtained with ultraviolet spectroscopic methods. The reproducibility and margin of error were tested with meat samples fortified with 11 PAHs and with samples of sunflower oil. The method was further applied to meat, smoked fish, yeast, and unrefined sunflower oil. All samples investigated contained more than 100 PAHs (characterized by mass spectrometry) of which only the main components were determined: phenanthrene, anthracene, fluorene, fluoranthene, pyrene, benzo(a)anthracene, chrysene, benzo(b)fluoranthene + benzo (j)fluoranthene + benzo(k) fluoranthene, benzo(e)pyrene, benzo(a)pyrene, perylene, dibenz(a,j)anthracene, dibenz(a,h)anthracene + indeno(1,2,3,-cd)pyrene, benzo(ghi)perylene, anthanthrene, and coronene. In contrast to other methods, the GLC profile analysis allows the recording of known and unknown PAH peaks simultaneously and also allows a compilation of all PAHs.     

Studying the Effects of Two Various Methods of Composting on the Degradation Levels of Polycyclic Aromatic Hydrocarbons (PAHs) in Sewage Sludge

The research comprised of studying the effect composting sewage sludge with sawdust and vermicomposting with earthworm Eisenia fetida has on the degradation of 16 polycyclic aromatic hydrocarbons (PAHs). Raw rural sewage sludge prior composting was more contaminated with PAHs than urban sewage sludge, in both cases exceeding EU cutoff limits of 6 mg/kg established for land application. Dibenzo[a,h]anthracene (DBahAnt), acenaphtylene (Acy) and indeno[1,2,3-c,d]pyrene (IPyr) were predominant in rural sewage sludge, whilst the urban sewage sludge contained the highest concentrations of benzo[b]fluoranthene (BbFl), benzo[k]fluoranthene (BkFl) and indeno[1,2,3-c,d]pyrene (IPyr). Thirty days of composting with sawdust has caused a significant reduction of 16 PAHs on average from 26.07 to 4.01 mg/kg (84.6%). During vermicomposting, total PAH concentration decreased on average from 15.5 to 2.37 mg/kg (84.7%). Vermicomposting caused full degradation of hydrocarbons containing 2 and 6 rings and significant reduction of PAHs with 3 aromatic rings (94.4%) as well as with 5 aromatic rings (83.2%). The lowest rate of degradation (64.4%) was observed for hydrocarbons with 4 aromatic rings such as fluoranthene, benzo(a)anthracene, chrysene and pyrene. On the other hand, the highest level of degradation was determined for PAHs with 2 rings (100%), 3 rings (88%) and 6 aromatic rings in the molecule (86.9%) after composting with sawdust. Acenaphthene and pyrene were found to be the most resistant to biodegradation during both composting methods.     

Polycyclic aromatic hydrocarbons in marine organisms from the Gulf of Naples, Tyrrhenian Sea

Polycyclic aromatic hydrocarbons (PAHs) were determined by an HPLC method with fluorescence detection in bivalves (Mitylus galloprovincialis), cephalopods (Todarodes sagittatus), crustaceans (Aristeus antennatus), and fish (Mullus surmeletus, Scomber scombrus, Micromesistius poutassou, and Merluccius merluccius) caught in the Gulf of Naples (Tyrrhenian Sea, Italy). Anthracene, fluoranthene, pyrene, benz(a)anthracene, chrysene, benzo(b)fluoranthene, and benzo(k)fluoranthene were detected, at different concentrations, in all of the examined marine organisms, whereas benzo(a)pyrene, dibenz(a,h)anthracene, benzo(g,h,i)perylene, and indeno(1,2,3-cd)pyrene were found only in Mediterranean mussels. Of mussels collected in winter 71.43% exceeded the maximum residual levels (MRL) fixed for the benzo(a)pyrene in European Regulation 208/2005/EC, whereas all samples collected in summer reported values lower than this limit. In comparison to the other marine organisms, the mussels showed the highest PAH concentrations (p < 0.01). Fish showed total PAH levels lower than those of cephalopods and, in particular, European hake showed the lowest values (6.06 ng/g of fresh weight).     

Characterization and Sources of PAHs and Potentially Toxic Metals in Urban Environments of Sevilla (Southern Spain)

The purpose of this study was to determine the degree of PAH contamination and the association of PAHs with metals in urban soil samples from Sevilla (Spain). Fifteen polycyclic aromatic hydrocarbons-PAHs (naphthalene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,h]anthracene, benzo[g,h,i]perylene, indeno[1,2,3-c,d]pyrene) and seven metals (Cd, Cr, Cu, Mn, Ni, Pb, Zn) have been evaluated in representative urban soil samples. Forty-one top soils (0–10 cm) under different land use (garden, roadside, riverbank and agricultural allotment) were selected. PAHs from soil samples were extracted by sonication using dichloromethane. The simultaneous quantification of 15 different PAH compounds were carried out by HPLC using multiple wavelength shift in the fluorescence detector. For qualitative analysis a photo diode-array detector was used. Metal (pseudo-total) analysis was carried out by digestion of the soils with aqua regia in microwave oven. The mean concentration of each PAH in urban soils of Sevilla showed a wide range, they are not considered highly contaminated. The results of the sum of 15 PAHs in Sevilla soils are in the range 89.5–4004.2 μg kg^?1, but there seems not to be a correlation between the concentration of PAHs and the land use. Of the 15 PAHs examined, phenanthrene, fluoranthene and pyrene were present at the highest concentrations, being the sum of these PAHs about 40% of the total content. Although metal content were not especially high in most soils, there are significant hints of moderate pollution in some particular spots. Such spots are mainly related with some gardens within the historic quarters of the city. The associations among metals and PAHs content in the soil samples was checked by principal components analysis (PCA). The largest values both for ‘urban’ metals (Pb, Cu and Zn) and for PAHs were mainly found in sites close to the historic quarters of the city in which a heavy traffic of motor vehicles is suffered from years.