添加不同营养助剂对石油污染土壤生物修复的影响

实验室条件下研究NPK复合肥、诺沃肥和腐殖酸三种物质不同配比添加对石油污染土壤的生物修复效果.在60d的修复实验中,定期取样测定土壤含油量、总异养菌数、石油烃降解菌数和脱氢酶活性,并采用PCR-DGGE技术研究修复过程中微生物多样性变化.结果表明,NPK肥的添加和NPK肥-诺沃肥-腐殖酸复合添加能够提高土壤中微生物的数量、微生物多样性和脱氢酶活性.在含油量为84600mg.kg-1的土壤中,添加营养助剂的处理60d后石油烃降解率为31.3%—39.5%,不添加营养助剂的石油烃降解率仅为3.5%.NPK肥-诺沃肥-腐殖酸复合添加对石油烃的降解率要高于NPK肥的单独添加(高8%),其原因可能是诺沃肥-腐殖酸能够有效提高土著微生物的活性,增加微生物的多样性,增强石油烃的降解.     

植物与微生物对石油污染土壤修复的影响

为了研究生物对石油污染土壤的修复效果,在从石油污染土壤中筛选石油降解微生物的基础上,采用盆栽试验,进一步研究了4种植物和筛选到的微生物对石油污染土壤修复的影响.选择中原油田地区的原油和潮土,采取人工污染方法,设计石油污染水平为15 g·kg-1.试验设置3类处理,即单独添加微生物、单独种植植物(分别为向日葵、狗牙根、棉花、高丹草)、微生物分别与4种植物(向日葵、狗牙根、棉花、高丹草)组合.结果表明:在石油污染土壤中单独添加石油降解微生物,120 d时,石油降解率达到67.0%;向日葵、狗牙根、棉花、高丹草对土壤中石油降解也具有一定效果,120 d时,石油降解率分别达到38.23%、36.57%、40.67%、38.67%;添加微生物和种植植物联合对石油降解能力大小顺序为棉花+微生物＞向日葵+微生物＞狗牙根+微生物＞高丹草+微生物.其中,棉花与微生物联合修复120 d可以使污染土壤石油降解率达到85.67%,在各种处理中对污染土壤中石油的降解效果最好.     

石油污染物的微生物降解研究

以炼油厂污水池底泥中分离的3株细菌和3株真菌为供试微生物,陕北黄土(0～20 cm)制备的土壤悬浮液为土壤微生物对照,灭菌培养基为非生物降解对照,研究不同组合微生物的生长动态及对石油烃的降解率.将不同组合的微生物接种到石油烃质量浓度为10000 mg·L-1的液体培养基中,29 ℃±1 ℃,摇床连续培养50 d.于0 ～72 h内取样,进行微生物生长动态检测;于5～50 d定期取样,考察培养基中石油烃降解率的动态变化.结果表明:单独外源菌降解效果优于混合外源菌和土著微生物,25 d后土著微生物生物降解率超过混合外源菌.外源细菌、外源真菌、混合外源菌和土著微生物50 d生物降解率分别为71.54 %,60.13 %,47.26 %和51.49 %.土壤细菌对石油污染物具有较强的生长适应性,外源细菌降解效果最好.     

多环芳烃污染土壤的微生物-紫花苜蓿联合修复效应

微生物-植物联合修复技术作为一种低耗高效的新型修复手段已经被广泛应用于有机污染土壤的修复领域并取得了较好的效果,新型生物资源的应用将推动该方法的进一步发展。本研究采用温室盆栽实验,以里氏木霉（Trichodermaressei FS10-C）、根瘤菌（Rhizobium meliloti）和紫花苜蓿（Medicago sativa L.）作为供试生物,设置添加灭活菌剂-无紫花苜蓿（CK）、添加灭活菌剂-种植紫花苜蓿（A）、接种木霉菌剂-种植紫花苜蓿（TA）、接种木霉菌根瘤菌复合菌剂-种植紫花苜蓿（TRA）4种处理,探究微生物-植物联合修复对多环芳烃（PAHs）污染土壤的生物修复效果及其微生态效应。结果表明,经过60 d的培养,微生物不仅促进了紫花苜蓿的生长,而且在紫花苜蓿的协同作用下进一步提高了土壤中PAHs降解率。TA处理中紫花苜蓿生物量增加了5.88%,而TRA处理进一步促进了紫花苜蓿的生长,其生物量增加了11.15%;A、TA和TRA处理下土壤中PAHs的降解率分别为17.02%、25.62%、32.93%,显著（p〈0.05）高于处理CK（5.67%）。此外,接种菌剂处理（TA、TRA）对土壤中高分子量PAHs具有更好的降解效果,A处理土壤中4-、5（＋6）环PAHs的降解率分别为18.13%、24.74%,TA处理为21.41%、28.34%,而TRA处理则为21.29%、30.11%。同时,紫花苜蓿能够通过其根际效应显著促进土壤微生物活性,相较于CK处理,A、TA、TRA处理土壤脱氢酶活性分别提高了33.20%、34.58%、32.65%,A、TA、TRA处理AWCD值和微生物群落多样性指数均显著（p〈0.05）高于CK。通过木霉、根瘤菌与紫花苜蓿联合作用不仅可以有效地降解土壤中的PAHs,而且能够恢复土壤微生物生态功能多样性和稳定性。因此,该方法是一种极具潜力的生物修复手段,具有广阔的市场应用前景。     

丛枝菌根真菌对紫花苜蓿与黑麦草修复多环芳烃污染土壤的影响

通过温室盆栽试验,研究接种土著与外源丛枝菌根(AM)真菌对紫花苜蓿与黑麦草修复多环芳烃(PAHs)污染土壤的影响.结果表明,接种外源AM真菌--苏格兰球囊霉(Glomus caledonium)36号能够显著提高紫花苜蓿和黑麦草的AM真菌侵染率并促进植物生长,而接种土著菌剂或土著菌剂与36号菌剂双接种对AM真菌侵染和植物生长无促进作用,甚至降低了黑麦草苗期的AM真菌侵染率.种植紫花苜蓿和黑麦草促进了土壤中PAHs的降解,这2种植物接种36号菌剂的处理60天时土壤PAHs降解率分别达42.3%和41.1%,说明36号菌剂可以显著提高植物修复效率,而接种土著菌剂对修复作用无显著影响,土著菌剂与36号菌剂双接种对紫花苜蓿的修复效果也无显著影响,但60天时显著提高黑麦草的修复效率.土壤中PAHs降解率与植物根系的AM真菌侵染率呈显著正相关关系(P<0.05),表明AM真菌侵染可以提高紫花苜蓿与黑麦草对PAHs污染土壤的修复效率.     

石油污染土壤的微生物修复及土壤微生物活性变化

为快速有效地测定石油污染土壤中功能性微生物的活性变化,分别以石油烃、正十六烷烃、多环芳烃为自定义碳源,应用Biolog法研究油污土壤生物修复过程中石油烃、烷烃、多环芳烃降解菌的代谢活性.结果显示,向油污土壤中投加混合降解菌群进行生物强化修复处理,可以有效去除土壤中的石油烃,修复13周土壤中石油烃去除率达到42.3%;生物刺激和自然修复对土壤石油烃的去除率分别为28.3%和20.5%.Biolog测定结果表明,生物强化法修复初期的土壤微生物群落对石油烃、烷烃两种碳源的代谢能力较强,而生物刺激法修复后期的土壤微生物群落对烷烃有较强的代谢能力;不同处理的土壤微生物群落比较偏好、利用率较高的碳源是石油烃,其次是烷烃,而对多环芳烃几乎不利用;土壤中石油烃、烷烃降解菌的活性越大,土壤微生物对石油烃的去除效率越高.上述研究结果说明,通过利用Biolog法测定土壤微生物活性变化可有效指示土壤中石油烃的去除效果.     

微生物对水-沉积物中苯并[a]芘-镉复合污染修复的研究

从广东汕头贵屿镇的沉积物中分离得到1株对PAHs有较好降解能力且有较强重金属耐受性的菌株（简称B4）,菌种鉴定表明该菌为氧化节杆菌属。以土著微生物、处理方式、沉积物、修复时间等为不同的影响因子对该菌修复水体/沉积物中苯并[a]芘（BaP）-镉（Cd）复合污染进行了初步研究。结果表明：该菌同时具有降解BaP和吸附Cd的性能;土著微生物对BaP有一定的降解能力,并能显著促进B4降解BaP,当体系中BaP质量浓度为1.00 mg.L-1时,降解率达到了68.3%;Cd的吸附却因土著微生物的存在发生一定程度的解吸。振荡培养对BaP的降解效果略优于静止培养修复,而静止修复却更有利于Cd的去除。沉积物促进体系Cd的吸附,却减弱了菌种对BaP的降解。在静止培养修复中,BaP的降解主要发生在7 d以前,降解率保持在35%左右,此时Cd的去除效果亦较好。     

基因工程菌在石油污染修复中的研究进展与前景

构建基因工程菌（genetically engineered microorganisms,GEMs）是石油污染生物修复的重要发展方向.目前,通过基因编辑、过表达和定向进化等手段改造微生物的石油污染物降解和调控途径,可以提高微生物的环境适应能力和污染物降解能力,用于石油污染物的生物降解和监测.本文概述了石油污染物降解基因工程菌的主要构建策略,包括选择和改造宿主菌、改造与优化石油污染物关键酶和代谢通路、开发微生物全细胞传感器和构建基因工程菌的自毁程序.此外,基因工程菌也可用于石油污染的酶修复、微生物菌群修复和细菌-植物联合修复.随着系统生物学和合成生物学在降解微生物中的应用,基因工程菌在石油污染修复中展现出良好的研究和应用前景.     

3种菌剂对水稻秸秆降解性能的影响

为寻求秸秆的高效快速处理途径,采用EM、VT1000、强兴3种菌剂作用于秸秆与果皮蔬菜混合垃圾按不同配比混合的物料,无需发酵,通过分析不同菌剂处理后秸秆混合物料的理化性质、降解性能和微生物种群结构变化,探讨不同菌剂对秸秆降解性能的影响。结果表明,3种菌剂均对物料的全碳、全氮、C/N具有降低作用,可提高速效磷和速效钾的含量,加快半纤维素的降解。EM对全碳、速效磷和C/N的作用效果较好,VT1000对全氮、速效钾和半纤维素的作用效果较好,且各菌剂于物料配比3?2时处理效果最佳,EM菌剂处理后物料全碳质量分数和C/N比值分别为8.19%和14.44,速效磷上升幅度为51.49%;VT1000菌剂处理后物料全氮下降幅度为55.24%,速效钾上升幅度为83.60%,半纤维素降解率为60.09%。采用高通量测序技术分析微生物群落结构对物料性能的影响,结果表明,不同菌剂处理后物料的优势细菌菌群为Proteobacteria(变形菌门)、Actinobacteria(放线菌门)、Bacteroidetes(拟杆菌门)、Acidobacteria(酸杆菌门)和Chlorobacteria(绿弯菌门),优势真菌菌群为Ascomycota(子囊菌门)、Zygomycota(接合菌门)、Basidiomycota(担子菌门)和Unclassified-k-Fungi(未经分类的真菌)。研究发现,优势菌群的相对丰度是影响秸秆性质变化的主要原因,EM菌剂处理的物料,优势菌群Unclassified-k-Fungi的相对丰度明显高于其他菌剂的处理,这直接影响物料的C/N变化;而VT1000菌剂处理的物料,优势菌群Acidobacteria相对丰度明显高于其他菌剂,其与物料中全氮和速效钾存在一定的相关性。该研究可为探索菌剂复配或研发新型菌剂用于秸秆的微生物降解奠定理论基础。     

水势与用氮量对土壤中石油降解的影响

研究了石油降解过程中，土壤有效氮的动态变化，不同用氮水平在两种土壤上对石油降解速率的影响，以及石油降解的土壤水势条件。结果表明，石油对土壤的污染没有影响土壤水势；在土壤水势为－１００￣－２０ｋＰａ范围内，水势的降低没有影响石油分解，而当土壤水势为－１０００ｋＰａ时，石油降解速率与对照相比下降了１０％￣３５％，在壤土上，肥料氮添加在碳（石油中碳）氮（肥料中氮）比为９０时，石油降解最快，过多施用肥料氮     

Bioremediation of oily sludge-contaminated soil by stimulating indigenous microbes

In situ bioremediation of oily sludge-contaminated soil by biostimulation of indigenous microbes through adding manure was conducted at the Shengli oilfield in northern China. After bioremediation for 360 days, total petroleum hydrocarbon (TPH) content was reduced by 58.2% in the treated plots compared with only 15.6% in the control plot. Moreover, bioremediation significantly improved the physicochemical properties of the soil in the treated plot. Soil microbial counts and community-level physiological profiling were also examined. Manure addition increased TPH degraders and polycyclic aromatic hydrocarbon (PAH) degraders in the contaminated soil by one to two orders of magnitude. The activity and biodiversity of soil microbial communities also increased markedly in the treated plot compared with that of the control. Finally, biotoxicity was used to evaluate the soils and a sharp increase in the EC50 of the soil after bioremediation was observed, indicating that bioremediation had reduced the toxicity of the soil.     

植物-微生物修复石油烃污染土壤与根际微生态环境变化

石油烃作为一类持久性难降解有机污染物对土壤环境质量产生严重的危害。以天津大港油田原油污染土壤中筛选出的耐低温高效石油烃降解菌为靶细胞,以小麦、紫花苜蓿作为供试植物,利用盆栽试验,对植物-外源菌协同修复体系中的脱氢酶活性和土壤微生物多样性进行研究,分析其变化及其与石油烃降解率的关系。结果表明植物-微生物协同修复对石油烃具有较好的降解能力,其中小麦-固定化外源菌组具有最高的降解率,石油烃含量从最初的30 600 mg獉kg-1下降为24 300 mg獉kg-1,降解率为20.6%,并且其试验后期石油烃的降解率最大,远远高于其他时期,表现出良好的修复潜力。外源菌投加的初始阶段会迅速提高脱氢酶活性,然而这种影响随着降解时间延长而逐渐减弱。初期脱氢酶活性与总石油烃的降解存在较好的相关性,脱氢酶活性可以在一定程度上表征土壤石油烃的降解情况。微生物多样性与总石油烃降解也存在一定的相关性。     

微生物修复土壤多环芳烃污染的研究进展

多环芳烃是一类具有致癌、致畸、致突变性质的持久性有机污染物,主要来源于煤、石油等燃料的不完全燃烧,易吸附于固体颗粒表面和有机腐殖质,化学结构稳定,能长期存在于自然环境,给人类健康和生态环境带来很大的危害。中国土壤多环芳烃污染严重,因此急需寻求有效的修复方法进行治理。在众多的多环芳烃污染修复方法中,微生物修复因其低成本、高效、污染少等优点成为研究热点。科学家们从自然界中分离出了多种细菌、真菌等具有降解多环芳烃能力的微生物,并对多环芳烃的降解机理进行了探索,结果表明,微生物在代谢活动过程中能够产生酶来实现对土壤中多环芳烃的降解。细菌主要通过产生双加氧酶来催化多环芳烃的加氧反应,而真菌可以通过分泌木质素降解酶系或单加氧酶来氧化多环芳烃。两种途径均是首先通过降低多环芳烃的稳定性,使之容易被进一步降解。目前,微生物修复技术正逐步应用于PAHs污染土壤的实地修复,且已取得一定成效。文章简要介绍了降解多环芳烃的微生物,对多环芳烃的微生物降解机制进行了综述,讨论了影响微生物修复过程的因素,列举了常见的微生物修复相关技术,展望了今后的研究趋势。     

固定化毛霉对工业和农田污染土壤中多环芳烃的降解和生物有效性评价

为了探讨微生物修复不同类型多环芳烃污染土壤的可行性,应用固定化毛霉对多环芳烃污染工业土壤及农田土壤进行微生物修复,用羟丙基-β-环糊精(HPCD)提取模拟评价多环芳烃的微生物可利用性,并分析多环芳烃微生物降解和生物可利用性的相关关系.焦化厂污染土壤中多环芳烃的30 d降解率为77.6%,沈抚灌区污染土壤中多环芳烃的30 d降解率为54.2%,焦化厂土壤和污灌区农田土壤中多环芳烃降解差异明显.焦化厂土壤和污灌区土壤中多环芳烃的30 d降解量和多环芳烃的环糊精可提取量具有相关性,各环数多环芳烃的环糊精可提取量变化解释了焦化厂和污灌区土壤中多环芳烃降解的差异机制,说明可用环糊精提取量预测微生物降解土壤多环芳烃的情况.     

Bioremediation of highly contaminated oilfield soil: Bioaugmentation for enhancing aromatic compounds removal

This study evaluated the effectiveness of different amendments--including a commercial NPK fertilizer, a humic substance （HS）, an organic industrial waste （NovoGro）, and a yeast-bacteria consortium--in the remediation of highly contaminated （up to 6% of total petroleum hydrocarbons） oilfield soils. The concentrations of hydrocarbon, soil toxicity, physicochemical properties of the soil, microbial population numbers, enzyme activities and microbial community structures were examined during the 90-d incubation. The results showed that the greatest degradation of total petroleum hydro- carbons （TPH） was observed with the biostimulation using mixture of NPK, HS and NovoGro, a treatment scheme that enhanced both dehydrogenase and lipase activities in soil. Introduction of exogenous hydrocarbon-degrading bacteria （in addition to biostimulation with NPK, HS and NovoGro） had negligible effect on the removal of TPH, which was likely due to the competition between exogenous and autochthonous microorganisms. None- theless, the addition of exogenous yeast-bacteria consor- tium significantly enhanced the removal of the aromatic fraction of the petroleum hydrocarbons, thus detoxifying the soil. The effect of bioaugmentation on the removal of more recalcitrant petroleum hydrocarbon fraction was likely due to the synergistic effect of bacteria and fungi.     

Influence of invasive plant Coreopsis grandiflora on functional diversity of soil microbial communities

Biological invasions are increasingly attracting the ecologists' attention. Invasive plants threaten the natural ecosystems not only by competing with the native plants, but also by altering the structure and function of soil microbial communities belowground. In this study, we studied the effects of the invasive plant Coreopsis grandiflora (C. grandiflora) on the functional diversity of soil microbial communities in Laoshan mountain in the province of Shandong, North of China. We sampled soil from plots that were invaded or not invaded by C. grandiflora. The functional diversity of microbial communities in the sampled soils was assessed by the Biolog procedure test. By the ANOVA analysis of average well color development (AWCD), Shannon index (H'), Shannon evenness (E), principle components analysis of the level physiological profiles (CLPP) and correlation analysis between the studied parameters, we found that the invasive species C. grandiflora enhanced the functional diversity of soil microbial communities where the habitat was invaded by the C. grandiflora. The study indicated thatthe successful invasive plants have profound effects on the function of soil microbial communities.     

Unexpected similar stability of soil microbial CO2 respiration in 20-year manured and in unmanured tropical soils

Soil respiration is one of the main CO₂ sources from terrestrial ecosystems. Soil respiration is therefore a major source of greenhouse gas. Knowledge of the impact of agronomic practices such as manuring on the stability, for example resistance and resilience, of heterotrophic C–CO₂ respiration to disturbance is scarce. Here, we studied the stability of soil microbial heterotrophic respiration of two tropical soils from plots annually enriched or not with manure applications during more than 20 years. Stability was quantified after heating soils artificially. We hypothesized that field manuring would change the stability of the microbial community. Additionally, the impact of both manured and unmanured soils to addition of an organic cocktail was assessed under controlled conditions in order to discriminate the metabolic capacity of the microbial community, and to link the metabolic capacity up with the microbial heterotrophic soil respiration. Our results show that total respiration was not significantly different in manured and unmanured pots. Moreover, contrary to our hypothesis, manure amendment did not affect the stability (resistance, resilience) of the microbial abundance or the basal metabolism, in our experimental conditions. By contrast, the diversity of the bacterial community in heated soils was different from that in unheated soils. After heating, surviving microorganisms showed different carbon utilization efficiency, manuring stimulating the growth of different resistant communities, that is, r-strategist or K-strategist. Microbial community of manured soils developed in the presence of the organic cocktail was less resistant to heating than microbial community of unmanured plots.     

In situ immobilisation remediation of cadmium in artificially contaminated soil: a chemical and ecotoxicological evaluation

A pot experiment was conducted to evaluate the effectiveness of sepiolite-induced immobilisation remediation of Cd contaminated soil. The results demonstrated that adding sepiolite significantly increased the soil pH and resulted in 35.1–66.0%, 30.3–48.9%, and 31.6–51.6% reduction in toxicity characteristic leaching procedure of Cd (TCLP-Cd), respectively, for the Cd levels of 1.25, 2.5, and 5?mg/kg compared with the no-sepiolite controls. These decreases in TCLP-Cd were associated with reductions in plant phyotoxicity and Cd absorption, and sepiolite-treated soils resulted in increases of 3.2–38.0%, 34.2–52.3%, and 8.4–51.5% in shoot biomass, respectively, and in decreases of 26.7–39.6%, 17.3–28.5%, and 6.1–21.8% in shoot Cd contents, respectively, under soil Cd concentration of 1.25, 2.5 and 5?mg/kg compared to the unamended soils. The greater microbial biomass and catalase and urease activities after applying sepiolite implied a certain degree of recovery in metabolic function recovery during soil remediation. These results demonstrated that the application of sepiolite not only was effective at reducing Cd bioavailability and the rate of Cd accumulation in plants, but also improved soil environmental quality.     

Changes in microbial community characteristics and soil organic matter with nitrogen additions in two tropical forests

Microbial communities and their associated enzyme activities affect the amount and chemical quality of carbon (C) in soils. Increasing nitrogen (N) deposition, particularly in N-rich tropical forests, is likely to change the composition and behavior of microbial communities and feed back on ecosystem structure and function. This study presents a novel assessment of mechanistic links between microbial responses to N deposition and shifts in soil organic matter (SOM) quality and quantity. We used phospholipid fatty acid (PLFA) analysis and microbial enzyme assays in soils to assess microbial community responses to long-term N additions in two distinct tropical rain forests. We used soil density fractionation and 13C nuclear magnetic resonance (NMR) spectroscopy to measure related changes in SOM pool sizes and chemical quality. Microbial biomass increased in response to N fertilization in both tropical forests and corresponded to declines in pools of low-density SOM. The chemical quality of this soil C pool reflected ecosystem-specific changes in microbial community composition. In the lower-elevation forest, there was an increase in gram-negative bacteria PLFA biomass, and there were significant losses of labile C chemical groups (O-alkyls). In contrast, the upper-elevation tropical forest had an increase in fungal PLFAs with N additions and declines in C groups associated with increased soil C storage (alkyls). The dynamics of microbial enzymatic activities with N addition provided a functional link between changes in microbial community structure and SOM chemistry. Ecosystem-specific changes in microbial community composition are likely to have far-reaching effects on soil carbon storage and cycling. This study indicates that microbial communities in N-rich tropical forests can be sensitive to added N, but we can expect significant variability in how ecosystem structure and function respond to N deposition among tropical forest types.     

有机酸与表面活性剂联合作用对土壤重金属的浸提效果研究

重金属污染土壤的原位淋洗修复既要实现对重金属的高效去除,还要尽量减少对土壤性质的破坏,这一点在农业污染土壤修复中尤为重要。以张士污灌区农田土壤为研究对象,利用振荡浸提技术筛选有机酸和表面活性剂组合,并确定了两者联合淋洗修复污染土壤的最佳配比。结果表明：有机酸（酒石酸、乙酸、柠檬酸和苹果酸）中的酒石酸浓度为0.5 mol.L-1和表面活性剂（SDBS、鼠李糖和皂素）中的皂素质量分数为0.7%时对土壤Cd、Pb、Zn的浸提效果较好;在酒石酸与皂素体积配比为1∶1时,对重金属Cd、Pb、Zn浸提效果最好,浸提率分别为87.62%、36.30%、20.67%;单一有机酸、表面活性剂或者有机酸与表面活性剂的混合溶液,对土壤重金属的浸提效果均为Cd〉Pb〉Zn。虽然有机酸与表面活性剂联合浸提效果略低于酒石酸浸提,但其弱酸性对土壤性质影响较小,在原位淋洗修复工程中有较好的应用前景。