生物泥浆反应器中多环芳烃微生物降解调控因子研究

多环芳烃 (PAHs)是一类普遍存在的环境中的优先有机污染物 ,在土壤中残留持久并难以降解 ,因此给环境和人体健康带来潜在危胁 [5] 。研究表明 ,微生物降解是去除 PAHs的主要途径 [6]。微生物能够以利用 PAHs作为碳源与能源和共代谢两种方式降解 PAHs[7] 。泥浆反应器作为生物修复技术之一 ,具有处理周期短 ,降解条件易于控制和处理效果好的特点[8,9] ,目前已成为一种重要的处理技术 ,在欧、美等发达国家 ,受到广泛重视。Grosser[10 ]等人发现 ,把微生物从污染地区分离培养后 ,接种到污染土壤中可大大提高芘的降解率。泥浆反应器中污染…     

海洋石油污染物的微生物降解与生物修复

石油是海洋环境的主要污染物 ,已经对海洋及近岸环境造成了严重的危害。微生物降解是海洋石油污染去除的主要途径。海洋石油污染物的微生物降解受石油组分与理化性质、环境条件以及微生物群落组成等多方面因素的制约 ,N和P营养的缺乏是海洋石油污染物生物降解的主要限制因子。在生物降解研究基础上发展起来的生物修复技术在海洋石油污染治理中发展潜力巨大 ,并且取得了一系列成果。介绍了海洋中石油污染物的来源、转化过程、降解机理、影响生物降解因素及生物修复技术等方面内容 ,强调了生物修复技术在治理海洋石油污染环境中的优势和重要性 ,指出目前生物修复技术存在的问题。     

低温微生物修复石油烃类污染土壤研究进展

耐冷菌、嗜冷菌等低温微生物广泛存在于极地、高山以及高纬度等土壤环境中,是石油烃类污染物在低温条件下降解与转化的重要微生物资源.利用低温微生物的独特优势,石油污染土壤的低温生物修复技术的研究成为当前热点领域.本文系统综述了低温石油烃降解菌的分类及冷适机制,低温微生物对不同类型石油烃组分的降解特征和降解机理,低温环境中接种降解菌、添加营养物质和表面活性剂等强化技术在石油污染土壤中生物修复的应用.以及微生物分子生物学技术在低温微生物降解石油烃的研究现状,为拓展我国石油污染土壤生物修复技术提供参考.     

实时荧光PCR在治理石油污染中的应用

石油污染已成为全球最严重的环境问题之一,运用生物学方法治理石油污染逐渐成为研究热点。从大港油田原油样品中筛选到12株能降解原油的菌株,分别对其降解特性进行了研究。其中,被命名为BS的菌株对原油的降解效果最好,降解率高达89.1%,经16S rDNA序列比对,与Bacillus subtilis BSn5的相似性达100%。通过生物信息学的方法,分析菌株B.subtilis BSn5的代谢途径,该菌株中有11个基因参与石油烃类化合物的降解过程。最后利用实时荧光PCR技术,测定在石油降解过程中起重要作用的烷基羟化酶基因(alkB)在菌株BS与标准菌株B.subtilis BSn5中的表达量,结果显示,该基因在菌株BS中的表达量较标准菌株B.subtilis BSn5高约2倍,这可能与菌株BS的生存环境有关。为检测和分析石油污染,筛选含有降解石油功能基因的微生物及构建工程菌株提供了新的手段。     

降解1,3-二甲基-2-咪唑烷酮细菌的分离及功能评价

[背景] 1,3-二甲基-2-咪唑烷酮（1,3-Dimethyl-2-Imidazolidinone,DMI）作为一种强极性非质子溶剂,在生产和应用过程的环境中有稳定残留问题,存在安全隐患。[目的] 分离筛选具有降解DMI能力的微生物菌株,为清除环境中残留的DMI提供优良的微生物菌种资源。[方法] 从DMI生产区域土壤采集样品分离DMI抗性微生物,采用形态学及分子生物学鉴定确定其分类地位,并对DMI降解能力进行测定。[结果] 分离到最高能够耐受5%（体积分数） DMI的微生物菌株,形态学及分子生物学鉴定初步表明获得的菌株DT-1和DT-2为贝莱斯芽孢杆菌（Bacillus velezensis）;全细胞及细胞提取液均具有降解DMI的能力;其中菌株DT-1及其细胞提取液对1%（体积分数） DMI的降解率分别达到48%和68%。[结论] 从DMI生产区域土壤中分离到具有DMI降解能力的芽孢杆菌,不但可为DMI污染的微生物治理提供优良微生物资源,而且扩展了人们对芽孢杆菌生物学功能的认识。     

高环多环芳烃降解菌的筛选及其降解特性

通过富集培养及平板升华法从本溪钢铁公司周边多环芳烃(PAHs)污染土壤中分离出7株PAHs降解菌。以芘和苯并[a]芘为底物进行摇瓶降解实验,结果表明:G1、G2和G3菌株对高环PAHs芘和苯并[a]芘均具有较强的降解能力。进一步研究此3株菌及混合菌对原状污染土壤中PAHs的降解能力,发现80 d时对总PAHs的降解顺序依次为:混合菌G2G1G3,其中混合菌对PAHs降解率较单菌分别提高了9.17%、11.49%和16.11%;4个处理对4~6环PAHs的降解率较对照组相比提高的倍数随着环数增加而增大;总PAHs的降解率与脱氢酶的活性呈正相关。电场影响G1、G2和G3菌株对PAHs降解,在1.0 V·cm~(-1)电场条件下,4环、5环及6环PAHs降解率较单纯微生物修复提高12.13%、13.35%和14.52%,说明3株菌具有较强的电场适应能力,可在高环PAHs污染土壤的电动-微生物修复中应用。形态学观察及16S rRNA序列比对分析表明,G1、G2、G3菌株分别为鞘氨醇单胞菌属(Sphingomonas sp.)、苍白杆菌属(Ochrobactrum sp.)和无色杆菌属(Achromobacter sp.)。     

菲污染土壤的微生物修复机制

菲(Phenanthrene)是存在于煤焦油中,含三个苯环的稠环芳烃。除了具有"三致"作用外,菲稳定的化学结构和高辛醇-水分配系数等特性,使其具备较强的抗降解能力,易在环境中富集,破坏土壤微生态结构,降低农作物品质,威胁人类健康。而且随着化石燃料的长期大量使用,受菲污染的土地面积也急速增加,给人类的健康及生产活动带来极大的威胁。因此,有效清除土壤中菲及其他多环芳烃污染物,净化环境,具有重要的现实意义。微生物降解作为治理菲污染的方法之一,具有高效、低成本、环境友好的特点,受到研究者的高度重视。本文从菲降解菌的种类、降解机理、分子机制、影响修复等因素及微生物与植物联合修复五个方面进行综述,为进一步利用环境微生物,开发高效菲降解菌,治理菲污染提供参考。     

土壤汞污染生物修复技术研究进展

汞作为常温下唯一的一种液态金属,在环境中分布十分广泛,极易造成土壤污染。生物修复是一种用于土壤汞污染治理的重要修复技术,该技术因具有修复成本低、绿色环保等优点而受到重视。对现有的土壤汞污染生物修复技术进行了评述,主要涉及植物修复技术、微生物修复技术和动物修复技术的研究和应用情况:植物修复技术主要研究汞在植物体内代谢机理和规律,微生物修复技术研究侧重于利用外源微生物降解土壤中汞,动物修复技术研究较少,目前仅见有关蚯蚓富集汞的报道。现代生物修复技术已经发展成为一门包括土壤化学、植物生物学、生态学、土壤微生物学、分析化学及分子生物学等多学科融合性技术,借助这些学科力量可以对植物提取汞根际微界面过程和植物体内微界面过程、微生物和动物汞富集机制有更深刻的认识。综述最后对目前土壤汞污染生物修复技术主要研究方向进行总结和归纳,明确了汞污染生物修复领域存在的几大问题以及研究方向。     

不同类型原油污染土壤生物修复技术研究

对不同类型原油污染土壤在实用规模的预制床上采用堆制技术进行生物修复 .通过投加肥料、菌剂、控制水分和pH ,可使微生物获得较好的生态环境 .当稀油、高凝油、特稠油和稠油污染的土壤中原油总量为 2 5 .8～ 77.2 g·kg-1土时 ,经过近 2个月的运行 ,石油总量的去除率可达 38.37%～ 5 6 .74 % .石油中芳烃、沥青和胶质混合物是制约石油快速降解的主要因素 .在处理过程中筛选出石油降解的优势菌株 ,其中有 6株真菌、6株细菌和 1株放线菌 .研究结果为石油污染土壤异位生物修复技术实用化提供了理论依据 .     

氟的危害及控制

环境中氟的危害是环境科学及卫生学界极为关注的问题。氟的过多吸收 ,对动植物及人体会产生危害[1 ,2 ] 。研究氟危害的表现特征及机理 ,目的是为了控制或减轻氟危害。综合评价氟的测定方法 ,在氟污染调查和环境质量评价中 ,有切实的实用价值。1　氟对植物及土壤微生物的毒害作用土壤氟污染对作物的危害是慢性积累的生理障碍过程。氟能抑制作物的新陈代谢、呼吸作用[3] 及光合作用[2 ] ,抑制新陈代谢过程中马来酸脱氢酶的活性[4] 。氟对作物的危害主要表现为干物质积累量少、产量降低、分蘖少、成穗率低、光合组织受损伤 ,出现叶尖坏死 ,叶…     

真菌对石油污染土壤的降解研究

利用微生物对石油污染土壤进行生物降解,具有操作简单,费用低廉,场地适用性强等特点。挑选了2种菌株,进行了室内油降解实验,在摇床实验油降解率：微生物真菌（Faserium．LK）（土著）和真菌（Phanerochaete．Chrysosprium）,在20d分别为41.2％和28．1％,真菌（Fusarium．LK）高于真菌（Phane－rochate．Chrysosprium）的降解率,而在培养箱石油污染土壤中,真菌（Fusarium．LK）（土著）和真菌（Phane－rochaete.Chrysosprium）,在50d分别为61.8％和66．1％,（Fusarium.LK）低于（Phanerochaete.Chrysosri-um）。     

Improvement of dry matter digestibility of water hyacinth by solid state fermentation using white rot fungi

Feeding value of water hyacinth biomass colonized by three species of white rot fungi during solid-state fermentation was investigated. All three organisms proved to be efficient degraders and enhanced dry matter digestibility. Loss of organic matter was maximum (23.6+/-0.1% dry wt) after 48 days by P. ostreatus. C. indica showed maximum cellulose degradation (18.5+/-0.1% dry wt) than other two fungi after 48 days of incubation. In all cases, an extensive removal of hemicellulose at the initial growth period and a delayed degradation of lignin were observed. Hemicellulolysis was maximum (46.3+/-0.1% dry wt) by C. indica, but delignification (14.2+/-0.2% dry wt) by P. sajor-caju after 48 days. The amount of reducing sugar in the degraded biomass decreased at early stages, but increased as degradation progressed in all three cases (maximum 1.1+/-0.05% dry wt after 48 days by C. indica). Soluble nitrogen content increased only during 16-32 days of incubation (highest 1.1+/-0.1% dry wt after 32 days by P. sajor-caju). Crude protein of the bioconverted biomass increased gradually up to 32 days but decreased thereafter (maximum 10.3+/-0.1% dry wt after 32 days by P. sajor - caju). Per cent change in in vitro dry matter digestibility of degraded substrates enhanced gradually after 8 days and reached maximum after 32 days but thereafter decreased (highest + 20.4+/-0.3% dry wt by P. sajor-caju). The results demonstrated the efficient degrading capacity of the test fungi and their potential use in conversion of water hyacinth biomass into mycoprotein-rich ruminant feed, more so by P. sajor-caju.     

植物法生物修复PAHs和矿物油污染土壤的调控研究

选择苜蓿草为供试植物,以污染物含量水平、专性细菌和真菌及有机肥为调控因子,进行了植物法生物修复多环芳烃(PAHs)和矿物油污染土壤的调控研究。结果表明,PAHs和矿物油的降解率与有机肥含量呈正相关,增加有机肥5%,可提高矿物油降解率17.6%～25.6%,PAHs降解率9%.在植物存在条件下,土壤微生物降解功能增强。多环芳烃总量的平均降解率比无植物对照土壤提高2.0%～4.7%.投加特性降解真菌可不同程度地提高土壤PAHs总量和矿物油的降解率。真菌对萤蒽、芘和苯(a)蒽/(艹屈)的降解有明显促进作用。而细菌能明显提高苊稀/芴、蒽和苯(a)萤蒽/苯(k)萤蒽的降解率。     

Comparative degradation of oomycete, ascomycete, and basidiomycete cell walls by mycoparasitic and biocontrol fungi

Fourteen fungi (primarily representing mycoparasitic and biocontrol fungi) were tested for their ability to grow on and degrade cell walls (CWs) of an oomycete (Pythium ultimum), ascomycete (Fusarium equisetii), and basidiomycete (Rhizoctonia solani), and their hydrolytic enzymes were characterized. Protein was detected in the cultural medium of eleven of the test isolates, and these fungi significantly degraded CWs over the 14-day duration of the experiment. In general, a greater level of CW degradation occurred for F. equisetii and P. ultimum than for R. solani. Fungi that degraded F. equisetii CWs were Coniothyrium minitans, Gliocladium roseum, Myrothecium verrucaria, Talaromyces flavus, and Trichoderma harzianum. Taxa degrading P ultimum CWs included Chaetomium globosum, Coniothyrium minitans, M. verrucaria, Seimatosporium sp., Talaromyces flavus, Trichoderma hamatum, Trichoderma harzianum, and Trichoderma viride. Production of extracellular protein was highly correlated with CW degradation. Considerable variation in the molecular weights of CW-degrading enzymes were detected among the test fungi and the CW substrates in zymogram electrophoresis. Multivariate analysis between CW degradation and hydrolysis of barley beta-glucan (beta1,3- and beta1,4-glucanases), laminarin (beta1,3- and beta1,6-glucanases), carboxymethyl cellulose (endo-beta1,4-glucanases), colloidal chitin (chitinases), and chitosan (chitosanases) was conducted. For F. equisetii CWs, the regression model accounted for 80% of the variability, and carboxymethyl cellulases acting together with beta-glucanases contributed an R2 of 0.52, whereas chitinases and beta-glucanases alone contributed an R2 of 0.11 and 0.12, respectively. Only 61% of the variability observed in the degradation of P. ultimum CWs was explained by the enzyme classes tested, and primarily beta-glucanases (R2 of 0.53) and carboxymethyl cellulases (R2 of 0.08) alone contributed to CW break down. Too few of the test fungi degraded R. solani CWs to perform multivariate analysis effectively. This study identified several fungi that degraded ascomyceteous and oomyceteous, and to a lesser extent, basidiomycetous CWs. An array of enzymes were implicated in CW degradation.     

表面活性剂TW-80对土壤中多环芳烃生物降解的影响

以表面活性剂TW80为供试物,进行了为期150d的实验研究,并分别在30、60和150d间隔采样监测PAHs降解率。结果表明,30d后,土壤中PAHs的降解率达90%,比对照提高约30%.60d后,浓度为10000mg·kg^-1表面活性剂的土壤和对照中,PAHs降解率从65.1%和60%迅速提高到93.8%和79.2%.其它处理中,PAHs的平均降解率仅比30d的结果提高4%.150d后,所有处理中PAHs的降解率均达到90%以上。可以认为,表面活性剂能提高PAHs的生物可利用性,加快PAHs的降解速率,从而减少污染暴露时间。但表面活性剂浓度过高可抑制微生物活性。研究还发现,TW80土壤中含有优势真菌。经鉴定为常见青霉、蠕形青霉、淡紫青霉和顶孢头孢霉。它们是土壤PAHs迅速降解的动因.     

土壤中毒死蜱和微生物相互作用的研究

实验结果表明,50、500、5000g·kg^-1的毒死蜱在灭菌土中的降解速度十分缓慢,60d后,仅降解20％左右。在施过毒死蜱的土壤中的降解速度非常迅速,60d后,50mg·kg^-1的降解率接近100％,500mg·kg^-1的降解率达到90％,5000g·kg^-1的降解率在80％左右。在未灭菌但未施过毒死蜱的土壤中的降解速度则介于二者之间．在各处理的土壤,50和500mg·kg^-1的毒死蜱在处理前期对真菌和细菌数量有一定的刺激作用,60d后,基本恢复到正常水平,而5000mg·kg^-1的毒死蜱则对真菌和细菌的数量有较强的抑制作用。60d后仍不能恢复,在同样处理条件下,施过毒死蜱土壤中的真菌和细菌数量并不比未施过毒死蜱土壤中的占优势,表明毒死蜱在施过药的土壤中降解速度的加快并不是由细菌和真菌的数量决定的,而是由它们的降解能力决定的。     

Biodegradation of petroleum hydrocarbons by filamentous fungi (Aspergillus ustus and Purpureocillium lilacinum) isolated from used engine oil contaminated soil

The use of microorganisms for remediation and restoration of hydrocarbons contaminated soils is an effective and economic solution. The current study aims to find out efficient telluric filamentous fungi to degrade petroleum hydrocarbons pollutants. Six fungal strains were isolated from used engine (UE) oil contaminated soil. Fungi were screened for their ability to degrade crude oil, diesel and UE oil using 2.6-dichlorophenol indophenol (DCPIP). Two isolates were selected, identified and registered at NCBI as Aspergillus ustus HM3.aaa and Purpureocillium lilacinum HM4.aaa. Fungi were tested for their tolerance to different concentration of petroleum oils using radial growth diameter assay. Hydrocarbons removal percentage was evaluated gravimetrically. The degradation kinetic of crude oil was studied at a time interval of 10 days. A.ustus was the most tolerant fungi to high concentration of petroleum oils in solid medium. Quantitative analysis showed that crude oil was the most degraded oil by both isolate; P. lilacinium and A. ustus removed 44.55% and 30.43% of crude oil, respectively. The two fungi were able to degrade, respectively, 27.66 and 21.27% of diesel and 14.39 and 16.00% of UE oil. As compared to the controls, these fungi accumulated high biomass in liquid medium with all petroleum oils. Likewise, crude oil removal rate constant (K) and half-lives (t_1/2) were 0.02 day⁻¹, 34.66 day and 0.015 day⁻¹, 46.21 day for P. lilacinium and A. ustus, respectively. The selected fungi appear interesting for petroleum oils biodegradation and their application for soil bioremediation require scale-up studies.     

Degradation of diesel oil in soil using a food waste composting process

We investigated the simultaneous degradation of diesel oil in soil and the organic matter in food waste by composting in 8 l reactors. Using a 0.5 l/min air flow rate, and 0.5-1% diesel oil concentrations together with 20% food waste, high composting temperatures (above 60°C) were attained due to the efficient degradation of the food waste. Petroleum hydrocarbons were degraded by 80% after only 15 days composting in the presence of food waste. In a 28 l reactor scale-up experiment using 1% oil, 20% food waste and 79% soil, removal efficiencies of petroleum hydrocarbons and food waste after 15 days were 79% and 77%, respectively.     

紫花苜蓿种子对逆境贮藏条件的反应

以陇东紫花苜蓿 (MedicagosativaL .cv .“Longdong”)种子为材料 ,在室温、35℃和 35℃ +10 %的种子含水量 (SMC) 3种贮藏、接种或不接种燕麦镰刀菌 (Fusariumavenaceum (Fr.)Sacc .)的条件下 ,1年贮藏期内对各逆境处理的种子每隔 6 0d进行 1次标准发芽试验 ,2 0℃恒温、第 10d统计种子的发芽率和死亡率 ,试验结束时计测种子幼苗的长度和感病率 ;在大田条件下观测各处理种子的出苗率 ,确定催腐 (CD)与各种贮藏条件下的苜蓿种带真菌种类和检出率 .结果表明 ,随着贮藏温度和种子含水量等逆境贮藏条件胁迫的加剧 ,苜蓿种带真菌检出率逐渐增高 ,从室温、35℃条件下的 10 %上升到CD +35℃ +10 %SMC条件下的 2 9% ;抗病性逐渐减弱 ,35℃ +10 %SMC条件下幼苗的感病率和种子死亡率显著 (P <0 .0 5 )高于室温和 35℃下的感病率和种子死亡率 ;室内种子发芽率和田间出苗率逐渐下降 ,35℃ +10 %SMC条件下的种子的发芽率和田间出苗率显著 (P <0 .0 5 )低于在室温和 35℃下的发芽率和田间出苗率 ;幼苗生长受到抑制 ,35℃ +10 %SMC条件下的苗长和根长显著 (P <0 .0 5 )低于在室温和 35℃下的幼苗长度 .随着贮藏时间的延长 ,种子真菌检出率和田间出苗率下降 ,幼苗感病率增加 .与未接种的对照相比 ,接种燕麦镰刀菌的种子     

Pyrene degradation by two fungi in a freshwater sediment and evaluation of fungal biomass by ergosterol content

Mucor racemosus var. sphaerosporus and Phialophora alba were investigated for their abilities to degrade pyrene in a freshwater sediment, with or without glucose supply as nutrient or carbon source, during 90 days. The ergosterol contents in sediment were quantified to estimate fungal biomass and to assess the correlation between fungal activity and biodegradation of pyrene. Results showed that, in an heterogeneous environment, these fungi presented different abilities to degrade pyrene. P. alba increased the degree of pyrene degradation by 9%, compared to the native micro-organisms, but a supply of glucose acted as an inhibitor to pyrene disappearance. M. racemosus var. sphaerosporus was not efficient at sediment bioremediation (with or without glucose added), because it reduced the rate of pyrene degradation by the native microflora. In any case, there was no increase of ergosterol in boxes during bioremediation experiments. In our experimental conditions, ergosterol content could not be correlated to pyrene degradation.