芦岭煤田微生物群落结构和生物成因气的产甲烷类型研究

【目的】揭示芦岭煤田微生物群落组成,并分析其潜在的产甲烷类型及产甲烷途径。【方法】采集芦岭煤田的煤层气样品和产出水样品,分别分析样品的地球化学性质特征;利用Illumina HiSeq高通量测序技术分析产出水中的微生物群落结构;采用添加不同底物的厌氧培养实验进一步证实芦岭煤田生物成因气的产甲烷类型。【结果】该地区煤层气为生物成因和热成因的混合成因气;古菌16S rRNA基因分析表明在产出水中含有乙酸营养型、氢营养型和甲基营养型的产甲烷菌。丰度较高的细菌具有降解煤中芳香族和纤维素衍生化合物的潜力。厌氧富集培养结果表明,添加乙酸盐、甲酸盐、H2+CO2为底物的矿井水样均有明显的甲烷产生。【结论】芦岭煤田具有丰富的生物多样性,该地区同时存在三种产甲烷类型。本研究为利用微生物技术提高煤层气的采收率,实现煤层气的可持续开采提供科学依据。     

哈密露天煤矿不同环境介质微生物群落特征分析

【背景】干旱区露天煤矿开采过程中产生的粉尘颗粒物加剧了土壤生态环境的恶化和矿区空气质量的下降,针对煤矿区土壤和粉尘颗粒物的微生物群落组成的研究鲜有报道。【目的】研究新疆哈密南湖乡露天煤矿土壤、粉尘及大气PM_2.5颗粒物中的微生物群落结构和多样性特征,并预测潜在的功能类群。【方法】采用高通量测序技术,对煤矿露天采坑区和电厂粉煤灰堆放区的土壤、粉尘及大气PM_2.5颗粒物的微生物真菌及细菌群落组成进行比对分析。【结果】矿区优势真菌类群来自子囊菌门(Ascomycota)和担子菌门(Basidiomycota),优势细菌类群来自变形菌门(Proteobacteria)和放线菌门(Actinobacteria)。真菌和细菌群落的丰富度及α多样性在整个矿区内无显著性差异,大气PM_2.5颗粒物的细菌群落生态位宽度显著大于露天采坑区和粉煤灰区。矿区内的土壤和PM_2.5颗粒物样本中均发现了一些丰度差异显著的功能类群,真菌特征功能类群为腐生营养型类群,细菌特征功能类群主要包括甲烷营养型类群、几丁质酶类细菌类群等。【结论】露天煤矿区粉尘可能对区域内土壤和PM_2.5颗粒物的微生物群落结构产生重要影响,具有煤组分降解功能的特定微生物类群可能是维持矿区土壤生态安全的重要微生物学机制之一。     

大柳塔长焰煤中灰分和无机矿物对生物产气的影响

【目的】以不同密度等级大柳塔长焰煤作为产气底物,前期驯化培养厌氧菌群进行生物模拟产气实验,研究不同密度等级煤中的灰分和无机矿物对生物产气的影响。【方法】利用小浮沉将大柳塔长焰煤分成不同密度等级的煤样,采用工业分析、XRD、XRF分析小浮沉处理得到煤样的理化性质,利用这些煤样进行生物产气模拟实验,以甲烷产量作为评价指标,分析不同密度等级煤样中灰分对产气的影响。最后,通过添加几种标准矿物方式比较了煤中无机矿物对生物产气的可能影响。【结果】不同密度等级煤样中灰分对产气量存在一般显著影响(P=0.035),且灰分与甲烷含量呈负相关关系,其灰分中的无机矿物如高岭土、菱铁矿、氧化亚铁镁等的积累对产气有抑制作用。不同矿物配比产气实验证实低含量的粘土矿物促进甲烷的生成,高含量的粘土矿物抑制产气。【结论】不同密度等级煤中的灰分对生物产气存在一般显著的影响,高灰分煤的产气量低,而低灰分煤的产气量高。     

贵州典型煤矸石堆场微生物群落结构及功能特征

【背景】煤矸石堆场用于堆放煤矿开采过程中产生的一种热值低、含重金属的固体废弃物,会产生大量的酸性废水,对堆场周边的生态环境造成严重影响。【目的】探究煤矸石堆场的微生物群落结构和功能特征。【方法】选择贵州省六枝特区典型煤矸石堆场作为研究对象,采集堆场表层土壤、矸石层土壤、废水浸出口沉积物和堆场下游河道沉积物,通过宏基因组学技术进行分析。【结果】细菌的优势菌门为变形菌门(Proteobacteria)和放线菌门(Actinobacteria),优势菌属为钩端螺菌属(Leptospirillum)和硫化杆菌属(Sulfobacillus);古菌的优势菌门为Candidatus＿Thermoplasmatota和泉古菌门(Crenarchaeota),优势菌属为热原体属(Thermoplasma)和金属球菌属(Metallosphaera)。不同采样点细菌和古菌的优势菌属存在显著差异,矸石层土壤和废水浸出口沉积物中的铁氧化细菌和硫氧化细菌比其他2个样点丰富。煤矸石堆场中微生物的碳、氮、硫代谢基因丰度较高,共检测到6条固碳途径、6条氮代谢途径和3条硫代谢途径。主要的固碳基因为ACAT和E2.2....     

2株草螺菌的鉴定及其对煤矸石胁迫下香根草的促生作用

【背景】植物-微生物联合修复技术在减轻环境胁迫作用、增强植物抗性、改善矿区景观等方面发挥重要作用,其中根际促生菌可与植物相互作用,促进植物生长,增强植物对非生物胁迫的耐受性。【目的】鉴定铜尾矿中分离得到的2株根际细菌,研究其对植物生长的促生特性,测定2株菌的接种对于煤矸石胁迫下香根草的影响。【方法】对铜尾矿中分离得到的2株菌进行16S rRNA基因序列鉴定和扫描电子显微镜观察,对菌株的促生特性进行测定,并将其分别接种于覆土5 cm的煤矸石和掺土10%的煤矸石中生长的香根草根际,测定2个月后香根草的理化指标和生理学指标。【结果】菌株P5-11和P5-19经鉴定均为草螺菌(Herbaspirillum),2株菌均具有固氮、溶磷、产吲哚乙酸(Indole-3-Acetic Acid,IAA)和产铁载体的特性,其中P5-19的产吲哚乙酸能力约为P5-11的2倍,具有较好的促生能力;2株菌均能提高香根草的株高、生物量、氮积累量和抗氧化酶活性,并降低丙二醛积累量。【结论】2株草螺菌均具有良好的促生特性,能够促进煤矸石胁迫下香根草的生长发育,这不仅为促生菌肥的研制提供了优良菌种,也为香根草在矿区生态恢复中的应用提供了参考价值。     

煤矿污染土壤降解苯并(a)芘微生物多样性及降解能力研究

以苯并(a)芘(50 mg/L)为唯一碳源,对新疆芦草沟煤矿开采区土壤微生物进行3代胁迫培养(每代60 d);采用PCR-DGGE方法了解不同污染程度土样中降解苯并(a)芘的微生物类群和多样性特点;利用高效液相色谱(HPLC)测定胁迫培养每代培养物混合菌群对苯并(a)芘的降解能力。PCR-DGGE结果显示:不同污染程度原始样品与苯并(a)芘胁迫培养第3代培养物的微生物香浓指数(H)、丰度(S)和均匀度(E)有所不同,其中重度污染培养物降解苯并(a)芘的微生物类群最丰富。对优势条带进行克隆,其主要归属于变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和放线菌门(Actinobacteria)。经HPLC检测发现重度污染样品中的群体微生物对苯并(a)芘的降解率明显高于轻度和中度污染样品,达到78.4%。研究表明新疆芦草沟煤矿开采区污染的土壤中可能蕴藏着降解苯并(a)芘的微生物资源。     

Solubilization of coal by biosurfactant derived from candida bombicola

Partial solubilization of North Dakota Beulah Zap lignite coal was demonstrated using crude biosurfactant preparations from glucose‐based stationary‐phase cultures of Candida bombicola. Results suggest that biosurfactant activity by this organism is a minor mechanism by which portions of coal can be biologically solubilized.     

Technological and economic aspects of coal biodesulfurisation

The sulfur found in coal is either part of the molecular coal structure (organically bound sulfur), is contained in minerals such as pyrite (FeS₂), or occurs in minor quantities in the form of sulfate and elemental sulfur. When pyrite crystals are finely distributed within the coal matrix, mechanical cleaning can only remove part of the pyrite. It can, however, be removed by microbial action requiring only mild conditions. The process involves simple equipment, almost no chemicals, but relatively long reaction times, and treatment of iron sulfate containing process water. Different process configurations are possibly depending on the coal particle size. Coal with particle sizes of less than 0.5 mm is preferably desulfurised in slurry reactors, while lump coal (> 0.5 mm) should be treated in heaps. Investment and operating costs are estimated for different process configurations on an industrial scale. Concerning the organically bound sulfur in coal there is up to now no promising biochemical pathway for the degradation and/or desulfurisation of such compounds.     

Colonization and degradation of oxidized bituminous and lignite coals by fungi

Summary APenicillium sp. previously shown to grow on lignite coals degraded an air-oxidized bituminous coal (Illinois #6) to a material that was more than 80% soluble in 0.5 N NaOH. Scanning electron microscopy of the oxidized Illinois #6 revealed colonization of the surface by thePenicillium sp., production of conidia, and erosion of the coal surface. The average molecular weight (MW) of Illinois #6 degraded by the fungus and base-solubilized was approximately 1000 Da. The average MW for base-solubilized Illinois #6 that was not exposed to the fungus was 6000 Da, suggesting solubilizing mechanisms other than base catalysis. A spectrophotometric assay to quantify the microbial conversion of biosolubilized coal was developed. Standard curves were constructed based on the absorbance at 450 nm of different quantities of microbe-solubilized coal. An acid precipitation step was necessary to remove medium and/or microbial metabolites from solubilized coal to prevent overestimation of the extent of coal biosolubilization. Furthermore, the absorption spectra for different coal products varied, necessitating construction of standard curves for individual coals.     

硫酸盐还原菌Desulfotomaculum reducens ZTS1厌氧降解昭通褐煤

[目的] 以云南昭通褐煤为研究对象,从矿井水富集产物中分离得到SRB菌株Desulfotomaculum reducens ZTS1厌氧降解褐煤,分析研究SRB菌株处理褐煤前后的物化性质变化。[方法] 采用工业和元素分析、XRD、FT-IR、SEM和低温液氮吸附法比较研究SRB厌氧降解褐煤前后矿物组成、有机官能团、表面形貌和孔隙结构变化。[结果] D.reducens ZTS1厌氧降解褐煤后灰分和挥发分略有降低,碳、氮和硫元素的含量降低,氧的含量增加,煤中硫代硫酸钠消失,FT-IR结果表明褐煤长链烃上的甲基和亚甲基团增多,游离羟基减少。煤表面粗糙度增加,小孔径的孔隙增多,孔径比表面积增大。[结论] D.reducens ZTS1可以厌氧降解昭通褐煤,厌氧降解后煤的物化性质发生变化。     

煤的预处理对高阶煤微生物溶解的影响

研究了煤的溶胀预处理和氧化预处理对微生物溶煤的影响,以及两种预处理方法处理过的煤样在固态培养基和液态培养基中的溶煤效果。考察了预处理方法、培养基选择、接种量等因素对溶煤作用的影响,得出了溶煤效果较好的适宜条件,并且对微生物溶煤的作用机理做了初步的探讨。     

复合微生物菌剂对水稻稻瘟病的生防效应

【背景】单一生防菌剂存在生防效果不够稳定、持效性差等不足,研究不同功能微生物的协同增效作用,开发复合微生物菌剂是控制植物病害的有效途径之一。【目的】探究不同功能微生物组合后对水稻稻瘟病的生防效应,开发高效生防水稻稻瘟病的复合微生物菌剂。【方法】将多株高效拮抗稻瘟病菌的链霉菌和细菌进行两两组合后与助剂复配,通过盆栽和大田试验,研究了复合微生物菌剂对水稻稻瘟病的生防效应。【结果】链霉菌Ahn75和解淀粉芽孢杆菌CWJ2菌株组合效果最好,与助剂复配后对水稻叶瘟和穗颈瘟的盆栽防效分别达到65.07%和63.00%,显著高于单一菌株Ahn75、CWJ2和其他菌剂组合的生物防效。同时,该复合菌剂能有效促进水稻植株生长,盆栽分蘖数和株高与对照相比分别提高93.33%和9.83%。而且,田间小区试验的结果也表明,该复合微生物菌剂的使用可以有效降低稻瘟病的发病,防效最高可达52.16%,与农药三环唑的防效(52.97%)相当。此外,菌株Ahn75和CWJ2分别对14种和16种稻瘟病病原菌生理小种的抑菌率超过50%,表明(Ahn75+CWJ2)复合菌剂对水稻稻瘟病具有广谱抗性。【结论】复合微生物菌剂(Ah...     

中国白酒发酵过程中的核心微生物群及其与环境因子的关系

【目的】揭示白酒酿造过程复杂微生物群落中的核心微生物群(core microbiota),定量分析核心微生物群的环境调控因素。【方法】通过高通量测序揭示发酵过程中的微生物群落结构,使用气相色谱-质谱联仪(GC-MS)测定发酵过程中的挥发性化合物。采用微生物群落与挥发性化合物轮廓关联分析获得风味代谢的功能微生物群(functional microbiota);通过微生物共现性网络分析,获得群落组成中的共现微生物群(co-occurring microbiota),两类微生物群的集合即为白酒酿造的核心微生物群。利用冗余分析(redundancy analysis)和蒙特卡洛置换检验(Monte Carlo permutation test)研究每个环境因素对该核心微生物群的影响。【结果】白酒发酵过程中的核心微生物群主要包含10个属,分别是Lactobacillus、Saccharomyces、Candida、Rhizopus、Saccharomycopsis、Pichia、Dipodascus、Bacillus、Thermoascus和Lactococcus。冗余分析和蒙特卡洛置换检验表明,化学因素(还原糖和乙醇)对核心微生物群的变化比物理因素(水分、温度和酸度)具有更加重要的影响作用,此外物理-化学因素的相互作用对核心微生物群的驱动也有很大的影响。【结论】本研究揭示了白酒发酵过程中的微生物群落组成和代谢物轮廓的变化规律及其二者之间的相关关系,确立了发酵过程中的核心微生物群并量化了影响核心微生物群变化的环境因素,为实现合成微生物组生产白酒及其定向调控奠定理论基础。     

Microbial desulfurization of solubilized coal

Microbial desulfurization of low rank coal by Rhodococcus rhodochrous IGTS8 was investigated using three different pretreated coal samples. Solubilized coal was desulfurized more efficiently than hard coal and more sulfur was extracted from biologically solubilized coal than from chemically solubilized coal. Microbial desulfurization combined with biological solubilization removed 75% of the total sufur while the microbial desulfurization combined with chemical solubilization removed 63%.     

Screening of microorganisms able to degrade low-rank coal in aerobic conditions: Potential coal biosolubilization mediators from coal to biochemicals

Coal is one of the major sources of energy, fuel, and other related chemicals. The processes to utilize coal for energy, fuel and other chemicals such as coal combustion, liquefaction, carbonization, and gasification pose a great threat to the environment by emitting toxic particles and CO₂ to the atmosphere. Thus, biological beneficiation of coal can be a good strategy to utilize coal with environmental sustainability. Here, we report the screening of microorganisms able to degrade or depolymerize coal. These host strains are potential candidates for the development of biological treatment process of coal. A total of 45 microbial strains were isolated from sludge enriched with coal and were identified based on 16S rRNA sequencing. Four strains of three genera, Cupriavidus sp., Pseudomonas sp., and Alcaligenes sp., were further characterized for their abilities to degrade coal. The degree of coal degradation was analyzed by measuring the increase in absorbance at 450 nm by UV spectroscopy. These microorganisms were also able to increase the pH of the culture media as a response to the acidic nature of coal. Laccase-like activity was also found in these strains when tested for RBBR dye degradation. Since biological degradation of coal through the use of microorganisms is a good alternative to chemical combustion of coal, microbial strains isolated in this study can be potential biological catalysts for coal conversion into valuable chemicals.     

Diversity of 16S rRNA and dioxygenase genes detected in coal‐tar‐contaminated site undergoing active bioremediation

Aims: In order to develop effective bioremediation strategies for polyaromatic hydrocarbons (PAHs) degradation, the composition and metabolic potential of microbial communities need to be better understood, especially in highly PAH contaminated sites in which little information on the cultivation‐independent communities is available. Methods and Results: Coal‐tar‐contaminated soil was collected, which consisted of 122·5 mg g^?1 total extractable PAH compounds. Biodegradation studies with this soil indicated the presence of microbial community that is capable of degrading the model PAH compounds viz naphthalene, phenanthrene and pyrene at 50 ppm each. PCR clone libraries were established from the DNA of the coal‐tar‐contaminated soil, targeting the 16S rRNA to characterize (i) the microbial communities, (ii) partial gene fragment encoding the Rieske iron sulfur center (α‐subunit) common to all PAH dioxygenase enzymes and (iii) β‐subunit of dioxygenase. Phylotypes related to Proteobacteria (Alpha‐, Epsilon‐ and Gammaproteobacteria), Acidobacteria, Actinobacteria, Firmicutes, Gemmatimonadetes and Deinococci were detected in 16S rRNA derived clone libraries. Many of the gene fragment sequences of α‐subunit and β‐subunit of dioxygenase obtained from the respective clone libraries fell into clades that are distinct from the reference dioxygenase gene sequences. Presence of consensus sequence of the Rieske type [2Fe‐2S] cluster binding site suggested that these gene fragments encode for α‐subunit of dioxygenase gene. Conclusions: Sequencing of the cloned libraries representing α‐subunit gene fragments (Rf1) and β‐subunit of dioxygenase showed the presence of hitherto unidentified dioxygenase in coal‐tar‐contaminated soil. Significance and Impact of the Study: The combination of the Rieske primers and bacterial community profiling represents a powerful tool for both assessing bioremediation potential and the exploration of novel dioxygenase genes in a contaminated environment.     

Comparison of coal solubilization by bacteria and fungi

Coal-solubilizing agents produced byTrametes versicolor, Phanerochaete chrysosporium, Aspergillus sp., a bacterial consortium, and a bacterial isolate,Arthrobacter sp., from that consortium were compared in terms of pH dependence, thermostability, molecular mass, mechanism of action, and product diversity. The thermostability and low molecular weights exhibited by the coal-solubilizing agents indicated a non-enzymatic mechanism of action. Competition studies using cupric copper indicated that coal solubilization by these agents involved metal chelation. Results demonstrated that oxalate could account for some but not all of the coal solubilization observed forT.versicolor andP.chrysosporium. The very low levels of oxalate detected inAspergillus sp. and the bacterial cultures indicated that oxalate is not an important factor in coal solubilization by these microbes. When subjected to gel permeation chromatography, the soluble coal products generated by each microbial coal-solubilizing agent yielded unique molecular mass profiles suggesting substantial product diversity. Such diversity increases the possibility of identifying potentially valuable compounds and extending the commercial utilization of coal.Abbreviations A₄₅₀, A₂₆₀ absorbances respectively at 450 nm and 260 nm - CSA coal-solubilizing agent - CSU coal-solubilizing unit - GPC gel permeation chromatography - MEA malt extract agar - PDA potato dextrose agar - SDA Sabouraud dextrose agar - SDB Sabouraud dextrose broth - SEM standard error of the mean - Tris tris(hydroxymethyl)aminomethane - TSA trypticase soy agar - TSB trypticase soy broth     

Solubilization of low-rank coal by Trichoderma atroviride: Evidence for the involvement of hydrolytic and oxidative enzymes by using 14C-labelled lignite

The deuteromycete Trichoderma atroviride is able to solubilize lignite in dependence on a given carbon source for growth. When cultivated on media containing glutamate, this mold excreted a set of different enzymes with hydrolytic activity. Addition of lignite to the growth media induced the synthesis of extracellular lignite-specific esterase activity but no evidence has been provided for its direct involvement in the process of lignite solubilization. Hence, the basic capability of T. atroviride enzymes to degrade a variety of ester and ether bonds at the surface or within the bulky lignite structure was tested using coal following its direct labelling with ¹⁴C-alkyl iodide. The participation of hydrolytic and oxidative enzymes in lignite degradation was assessed by measuring the release of ¹⁴C radioactivity from selectively alkylated carboxylic and phenolic OH groups. T. atroviride cleaved both carboxylic esters using esterases and the phenolic ether bonds by using oxidative enzymes, most likely laccases. Journal of Industrial Microbiology & Biotechnology (2002) 28, 207–212 DOI: 10.1038/sj/jim/7000232 Received 05 July 2001/ Accepted in revised form 08 December 2001     

内源微生物驱油及其对油藏微生物活动的影响

【目的】新疆油田六中区为典型水驱普通稠油油藏，水驱效果较差，油藏具有丰富的内源微生物，本研究通过分析内源微生物驱油对油藏微生物活动的影响，确定内源微生物驱油技术在该类油藏的应用潜力。【方法】采用高通量测序及分析化学技术，系统研究实施内源微生物驱油技术后油藏细菌群落结构组成、细菌总数和功能菌群的浓度以及采出液的流体性质，总结内源微生物驱油对油藏微生物活动的影响。【结果】现场试验注入激活剂和空气后，内源微生物被显著激活，细菌群落结构发生明显变化，细菌总数及功能菌群浓度普遍提高了2–3个数量级；各种内源微生物代谢活动显著增强，与地层流体相互作用后，原油明显被乳化，最终石油采收率提高5.2%。【结论】对于内源微生物较为丰富的水驱普通稠油油藏，内源微生物驱油技术对油藏微生物活动的影响显著，具有显著的技术优势和较大的应用潜力，微生物群落结构、功能菌群浓度及其相关代谢产物可以作为评价内源微生物驱油现场激活效果的重要指标，为其他内源微生物驱油现场试验提供技术参考。     

过氧化氢预处理对褐煤物化性质及生物产气的影响

【目的】研究过氧化氢预处理对褐煤物化性质及生物产气的影响。【方法】以胜利5号褐煤为研究对象,利用正交试验对过氧化氢预处理褐煤条件进行优化,在最优条件下处理褐煤得到处理后的残煤和处理液,通过X射线衍射分析(X-ray diffraction, XRD)、扫描电镜分析(scanning electron microscopy, SEM)、比表面积分析及孔隙分析(brunauer-emmett-teller, BET)、气相色谱-质谱分析(GC-MS)、高效液相色谱分析(HPLC)等方法对原煤、残煤和处理液的物化性质进行比较分析。【结果】经过氧化氢预处理,褐煤的最优条件为过氧化氢浓度5.0%、预处理时间20 d、液固比30:1,处理液中总有机碳含量为105 mg/L。在最优条件下,过氧化氢处理后残煤表面裂痕、凹陷增多,表面结构变得松散;煤的芳香面网间距增加,芳环结构更加疏松,晶核结构变小;孔隙度和比表面积均增大。处理后残煤中的固定碳、C元素和镜质组的相对含量降低,而灰分、挥发分、O和H元素及惰质组含量增加,残煤中O=C-O、C=C、C=O官能团含量增加,而N-H、C-H官能团含量则减少。生物产气结果表明反应液和残煤产气量均低于原煤,分别减少了39.13%和94.46%。过氧化氢预处理主要作用于煤中镜质组,使其有机碳溶解,煤中大分子结构的官能团发生变化,改变煤的芳环结构,在氧化作用下煤结构中的小分子溶解进入处理液。处理液中有机物以短链脂肪酸为主。经生物产气后,反应液中小分子酸以及有机物种类减少,被微生物利用产气。而各产气试验组中优势菌门及优势菌属的菌群丰度呈现出显著差异,古菌中原煤产气组盐杆菌门(Halobacteriota)为优势菌门,甲烷八叠球菌属(Methanosarcina)为优势菌属;反应液产气组热变形菌(Thermoprotei)为优势菌门,深古菌属(Bathyarchaeia)为优势菌属;细菌中原煤产气组放线菌门(Actinomycetota)为优势菌门,Gaiellales为优势菌属;反应液产气试验组假单胞菌门(Pseudomonadota)为优势菌门,代尔夫特菌属(Delftia)为优势菌属。【结论】煤溶解有机碳可以被微生物利用产气,但是煤中有机组分的过氧化脱除导致生物产气量减少。