页岩气开采工艺的微生物分子生态学研究进展

页岩气因其储量丰富及绿色清洁等特点已成为能源领域的研究热点。微生物对页岩气的形成和开采过程有一定影响,研究与页岩气相关的微生物分子生态,将有助于解析页岩气成因,改进页岩气生产设备及产出水等的管理。本文阐述了国内外对微生物在页岩气形成及开采方面的研究进展,包括页岩气成因类型,微生物产甲烷途径及环境对产甲烷途径的影响,钻井液和水力压裂液对微生物群落结构的影响及微生物对生产设备的影响,指出深层页岩气井生产设备中微生物分子生态和商业开采对地下微生物的影响可作为今后的关注重点,旨在为我国页岩气产业的可持续发展提供参考。     

页岩气压裂返排液及排放废液的研究现状及微藻资源化处理应用前景综述

在页岩气勘探开发的诸多环境问题中, 页岩气压裂返排液及排放废液污染问题表现尤为突出, 压裂返排液及排放 废液组分的复杂性及性质的独特性决定了其处理难度大、费用高, 被普遍认为是最难处理的工业污水之一, 如何合理处置页岩气开发中产生的大量返排液及排放废液已成为页岩气规模化开发的重要瓶颈问题之一。文章综述了页岩气压裂返排液及排放废液的环境影响与风险及处理现状, 并针对压裂返排液及排放废液的水质特点, 讨论了相应的资源化应用前景。     

页岩气开发对植被和土壤的影响研究进展

页岩气开发和生产过程影响水资源与水质、土地利用与植被覆盖、土壤侵蚀与土壤质量。综述了页岩气开发和生产过程可能存在的生态环境影响,并重点评价了国际上不同区域页岩气开发和生产对土地利用和植被覆被变化、景观破碎化的影响,以及对土壤侵蚀和土壤质量的影响。研究发现页岩气开发平台、运输道路和管线占用农田、牧场、森林,造成不同程度的景观破碎化,在坡地开发页岩气会导致土壤侵蚀与沉积风险增大。截止2015年末,在我国重庆涪陵焦石坝页岩气产建区55.8%的面积(146.56 km~2)存在土壤侵蚀和石漠化生态风险。我国页岩气开发区水基钻屑固化填埋未对周边土壤造成污染。建议页岩气开发设计应考虑占地、景观破碎化的影响,并及时开展页岩气开发暂时占地的复垦工作。     

涪陵焦石坝页岩气开采区土地损毁的生态风险评价

页岩气在勘探、开采、集输过程中对周边生态系统会产生直接或间接影响。通过构建页岩气开采土地损毁生态风险因果链识别井场、运输道路及集输管线等风险源,基于最小阻力模型定量分析土地损毁的生态累积影响;选取了植被覆盖度、生态服务价值及土壤肥力等因子表征区域生态重要性,土壤侵蚀度、石漠化敏感性及水环境敏感性等表征生态脆弱性,综合评价区域生态敏感度,并以此为风险受体,实现涪陵焦石坝页岩气开发区的生态风险评价。2012—2015年末,涪陵焦石坝页岩气产建区钻井数量快速增加,分布广,页岩气开发对区域生态累积影响扩大。2015年末,区域一半以上面积为中、高生态风险区(146.56km~2,55.8%),主要分布于南部乌江河谷及北部低山区,前者水环境敏感度高,后者岩溶发育度高,土壤侵蚀度高,石漠化敏感;该区域内大规模页岩气开发将面临水环境污染、生境破坏、土壤退化、石漠化加重及生物多样性减少等生态风险,是生态环境管理及风险防范的重点方面。研究结果可为区域生态安全建设提供科学的参考。     

微生物勘探技术在能源勘探中的应用

<正>目前流行的石油、天然气和铀矿开采技术,主要是基于地质科学和地球物理科学的勘探技术。而基于微生物技术的烃类气体勘探技术则较为少用。烃类气体勘探技术的基础是烃类气体能够通过油气藏储层结构的土壤缝隙向地表迁移。早期的石油和天然气勘探就是通过甲烷或原油的地表面微渗漏发     

祁连山天然气水合物赋存区钻孔细菌多样性

分析了青海省祁连山冻土区天然气水合物赋存区DK-6钻孔的4个样品,对岩心样品进行处理提取微生物总基因组,采用PCR构建了细菌16S rDNA基因文库,4个文库包括44个OTU,其中有厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、变形杆菌门(Proteobacteria)(包括α-、β-和y-变形杆菌亚群)、放线菌纲(Actinobacteria)和异常球菌-栖热菌门(Deinococcus-Thermus)5类,煤、泥岩、粉砂岩等不同岩性的微生物群落之间显示出较大的组成差异,优势菌不同.细菌菌群多样性随采集点地质环境不同而有较明显的变化,天然气水合物含量、水含量、有机质含量等环境因素对冻土区天然气水合物赋存区中细菌菌群有一定的影响.4个样品中存在的微生物大部分可以代谢有机烃类,在天然气水合物环境的特殊条件下,外界环境因素制约了微生物的种类.     

地质封存二氧化碳与深地微生物相互作用研究进展

地质封存将工业和能源相关领域生产活动产生的二氧化碳（CO₂）进行捕集并注入到深部地下岩石构造中,以实现长期储存的目标,是降低温室气体排放、实现CO₂长期封存的重要可行性手段之一。向深部地下地质构造中注入大量CO₂会导致深地环境发生显著变化,进而引起原生微生物活性及群落结构发生明显改变。因此,地质封存CO₂能够直接或间接影响深地微生物驱动的生物地球化学过程。同时,微生物在短期和长期的超临界CO₂（scCO₂）胁迫作用下,也会通过不同的适应性进化方式影响CO₂在地下环境中的迁移、转化和赋存形态。本文介绍了国内外二氧化碳捕获与封存发展现状以及地质封存CO₂影响条件下的scCO₂-水-微生物-矿物的相互作用领域的最新科研进展,并展望了利用深地微生物强化CO₂固定以及将其转化为高附加值产物的潜力。     

深松对乌拉尔甘草根际土壤养分以及微生物群落功能多样性的影响

通过大田试验和室内分析相结合,研究了深松对乌拉尔甘草根际土壤养分和微生物群落功能多样性的影响,以期为乌拉尔甘草人工种植地土壤耕作措施优化和土壤环境改良提供依据。结果表明,与未深松(常规耕作)处理相比,深松处理对乌拉尔甘草根际土壤0—20 cm耕层土壤养分含量无显著性影响,可显著提高乌拉尔甘草根际土壤20—40 cm耕层有机质、全氮、全磷和全钾的含量,分别提高了60.8%、65.3%、48.9%和86.8%;明显增加了0—20 cm和20—40 cm耕层细菌、真菌和放线菌的数量(P0.05),3种类型的微生物数量均呈现出上层大于下层,深松大于未深松的变化趋势。在156 h的微生物温育期内,深松处理下不同土层的平均颜色变化率(AWCD)均显著高于未深松处理,并显著提高了AWCD的利用率(72 h,P0.05),较未深松分别提高了35.5%和130.8%。与未深松处理相比,深松处理显著提高了土壤微生物的多样性指数(H、S、D)。主成分分析(PCA)表明,深松优化了乌拉尔甘草根际土壤微生物的群落组成;聚合物、羧酸类化合物、氨基酸和碳水化合物是深松处理下根际土壤微生物利用的主要碳源。总而言之,深松处理显著提高乌拉尔甘草根际土壤养分含量、微生物数量和微生物多样性指数,改变了微生物群落功能多样性,造成这种差异的主要原因可能是深松改变了土壤耕层结构,改善了微生物的生存环境。因此,深松对乌拉尔甘草人工种植地土壤质量的改良有积极作用。     

石油烃的厌氧生物降解对油藏残余油气化开采的启示

利用微生物将油藏中难以动用的原油就地转化为甲烷,以天然气的形式开采、或作为战略资源就地储备,从而大幅度提高油气资源的利用率,是当前国际上研究的前沿课题。本文综述了石油烃厌氧生物降解转化为甲烷的菌群结构、反应热力学和反应动力学等基础科学问题的最新研究进展,讨论了油藏残余油气化开采技术的可行性及开发潜力,提出了该技术进一步研究的方向。     

农用化学品污染对土壤微生物群落DNA序列多样性影响研究

采用ＲＡＰＤ分子遗传标记技术研究了农用化学品不同使用环境下的４种土壤微生物群落ＤＮＡ序列多样性的变化。结果表明,４种土壤微生物群落ＤＮＡ序列在其丰富度、多样性指数、均匀度等方面均存在差异;农用化学品的使用会对土壤微生物群落在ＤＮＡ分子水平上的多样性产生影响;而冰同的农用化学品对土壤微生物群落ＤＮＡ序列多样性影响各不相同：化肥污染会引起某些土壤微生物的富集和一些微生物物种的丧失;农药杂会引起土壤微生     

Bacterial communities associated with hydraulic fracturing fluids in thermogenic natural gas wells in North Central Texas, USA

Hydraulic fracturing is used to increase the permeability of shale gas formations and involves pumping large volumes of fluids into these formations. A portion of the frac fluid remains in the formation after the fracturing process is complete, which could potentially contribute to deleterious microbially induced processes in natural gas wells. Here, we report on the geochemical and microbiological properties of frac and flowback waters from two newly drilled natural gas wells in the Barnett Shale in North Central Texas. Most probable number studies showed that biocide treatments did not kill all the bacteria in the fracturing fluids. Pyrosequencing-based 16S rRNA diversity analyses indicated that the microbial communities in the flowback waters were less diverse and completely distinct from the communities in frac waters. These differences in frac and flowback water communities appeared to reflect changes in the geochemistry of fracturing fluids that occurred during the frac process. The flowback communities also appeared well adapted to survive biocide treatments and the anoxic conditions and high temperatures encountered in the Barnett Shale.     

A structured hazard identification method of human error for shale gas fracturing operation

Shale gas fracturing is a complex system of continuous operation. If human errors occur, it will cause a chain reaction, from abnormal events to fracturing accidents, and even lead to abandonment of shale gas wells. The process of shale gas fracturing has many production stages that are complex and the consequence of any error is serious. The human error modes in shale gas fracturing process are mutative. Therefore, human error should be studied in a systematic way, and in a hybrid framework, that is, whole integration of identification, prioritization, reasoning, and control. A new structured identification method of human error in a hybrid framework for shale gas fracturing operation is presented in the article. First, human error is structurally identified based on the human HAZOP method. Second, fuzzy VIKOR method is applied to comprehensive prioritization. Finally, 4M element theory is used to analyze the human error and control its evolution. The method improves the consistency of the identification results through the standard identification step and the identification criterion. Results from a study of feed-flow process indicate that 34 kinds of human errors can be identified, and high-probability errors occur in the behavior of implementation and observation.     

Geological gas-storage shapes deep life

Around the world, several dozen deep sedimentary aquifers are being used for storage of natural gas. Ad hoc studies of the microbial ecology of some of them have suggested that sulfate reducing and methanogenic microorganisms play a key role in how these aquifers' communities function. Here, we investigate the influence of gas storage on these two metabolic groups by using high-throughput sequencing and show the importance of sulfate-reducing Desulfotomaculum and a new monophyletic methanogenic group. Aquifer microbial diversity was significantly related to the geological level. The distance to the stored natural gas affects the ratio of sulfate-reducing Firmicutes to deltaproteobacteria. In only one aquifer, the methanogenic archaea dominate the sulfate-reducers. This aquifer was used to store town gas (containing at least 50% H₂) around 50 years ago. The observed decrease of sulfates in this aquifer could be related to stimulation of subsurface sulfate-reducers. These results suggest that the composition of the microbial communities is impacted by decades old transient gas storage activity. The tremendous stability of these gas-impacted deep subsurface microbial ecosystems suggests that in situ biotic methanation projects in geological reservoirs may be sustainable over time.     

Geochemical Influence on Diversity and Microbial Processes in High Temperature Oil Reservoirs

The diversity of thermophilic microbial assemblages detected within two neighboring high temperature petroleum formations was shown to closely parallel the different geochemical regimes existing in each. A high percentage of archaeal 16S rRNA gene sequences, related to thermophilic aceticlastic and hydrogenotrophic methanogens, were detected in the natural gas producing Rincon Formation. In contrast, rRNA gene libraries from the highly sulfidogenic Monterey Formation contained primarily sulfur-utilizing and fermentative archaea and bacteria. In addition to the variations in microbial community structure, microbial activities measured in microcosm experiments using high temperature production fluids from oil-bearing formations also demonstrated fundamental differences in the terminal respiratory and redox processes. Provided with the same suite of basic energy substrates, production fluids from the South Elwood Rincon Formation actively generated methane, while thermophilic microflora within the Monterey production fluids were dominated by hydrogen sulfide producing microorganisms. In both cases, molecular hydrogen appeared to play a central role in the stimulation of carbon and sulfur cycling in these systems. In methanogenic production fluids, the addition of sulfur compounds induced a rapid shift in the terminal electron accepting process, stimulating hydrogen sulfide formation and illustrating the metabolic versatility of the subsurface thermophilic assemblage. The high similarity between microbial community structure and activity corresponding with the prevalent geochemical conditions observed in deep subsurface petroleum reservoirs suggests that the resident microflora have adapted to the subsurface physicochemical conditions and may actively influence the geochemical environment in situ.     

Influence of the drilling mud formulation process on the bacterial communities in thermogenic natural gas wells of the Barnett Shale

The Barnett Shale in north central Texas contains natural gas generated by high temperatures (120 to 150°C) during the Mississippian Period (300 to 350 million years ago). In spite of the thermogenic origin of this gas, biogenic sulfide production and microbiologically induced corrosion have been observed at several natural gas wells in this formation. It was hypothesized that microorganisms in drilling muds were responsible for these deleterious effects. Here we collected drilling water and drilling mud samples from seven wells in the Barnett Shale during the drilling process. Using quantitative real-time PCR and microbial enumerations, we show that the addition of mud components to drilling water increased total bacterial numbers, as well as the numbers of culturable aerobic heterotrophs, acid producers, and sulfate reducers. The addition of sterile drilling muds to microcosms that contained drilling water stimulated sulfide production. Pyrosequencing-based phylogenetic surveys of the microbial communities in drilling waters and drilling muds showed a marked transition from typical freshwater communities to less diverse communities dominated by Firmicutes and Gammaproteobacteria. The community shifts observed reflected changes in temperature, pH, oxygen availability, and concentrations of sulfate, sulfonate, and carbon additives associated with the mud formulation process. Finally, several of the phylotypes observed in drilling muds belonged to lineages that were thought to be indigenous to marine and terrestrial fossil fuel formations. Our results suggest a possible alternative exogenous origin of such phylotypes via enrichment and introduction to oil and natural gas reservoirs during the drilling process.     

Feast and famine--microbial life in the deep-sea bed

The seabed is a diverse environment that ranges from the desert-like deep seafloor to the rich oases that are present at seeps, vents, and food falls such as whales, wood or kelp. As well as the sedimentation of organic material from above, geological processes transport chemical energy--hydrogen, methane, hydrogen sulphide and iron--to the seafloor from the subsurface below, which provides a significant proportion of the deep-sea energy. At the sites on the seafloor where chemical energy is delivered, rich and diverse microbial communities thrive. However, most subsurface microorganisms live in conditions of extreme energy limitation, with mean generation times of up to thousands of years. Even in the most remote subsurface habitats, temperature rather than energy seems to set the ultimate limit for life, and in the deep biosphere, where energy is most depleted, life might even be based on the cleavage of water by natural radioisotopes. Here, we review microbial biodiversity and function in these intriguing environments.     

Bioremediation of groundwater pollution.

Significant progress has been made in the past year towards an understanding of the microbial processes in subsurface environments that may allow natural microbial populations to be employed for bioremediation of groundwater pollution. Among the highlights were: the discovery of several previously unknown xenobiotic-degrading abilities in groundwater microorganisms; progress in using the unique abilities of methanotrophs to oxidize halogenated solvents; and characterizations of microbial populations from subsurface soils.     

Microbial activities in soil near natural gas leaks

Summary From the present experiments it may be concluded that in the surroundings of natural gas leaks, methane, ethane and possibly some other components of the natural gas are oxidized by microbial activities as long as oxygen is available. This is demonstrated by an increased oxygen consumption and carbon dioxide production, as well as by increased numbers of different types of bacteria. The resulting deficiency of oxygen, the excess of carbon dioxide, and perhaps the formation of inhibitory amounts of ethylene, are considered to be mainly responsible for the death of trees near natural gas leaks. Also the long period of time needed by the soil to recover, may be due to prolonged microbial activities, as well as to the presence of e. g. ethylene.The present experiments suggest that especially methane-oxidizing bacteria of the Methylosinus trichosporium type were present in predominating numbers and consequently have mainly been responsible for the increased oxygen consumption. However, some fungi oxidizing components of natural gas, including methane and ethane may also have contributed to the increased microbial activities in the soil. The same will be true of a possible secondary microflora on products derived from microorganisms oxidizing natural gas components.     

Desulfotomaculum and Methanobacterium spp. dominate a 4- to 5-kilometer-deep fault

Alkaline, sulfidic, 54 to 60 degrees C, 4 to 53 million-year-old meteoric water emanating from a borehole intersecting quartzite-hosted fractures >3.3 km beneath the surface supported a microbial community dominated by a bacterial species affiliated with Desulfotomaculum spp. and an archaeal species related to Methanobacterium spp. The geochemical homogeneity over the 650-m length of the borehole, the lack of dividing cells, and the absence of these microorganisms in mine service water support an indigenous origin for the microbial community. The coexistence of these two microorganisms is consistent with a limiting flux of inorganic carbon and SO4(2-) in the presence of high pH, high concentrations of H2 and CH4, and minimal free energy for autotrophic methanogenesis. Sulfide isotopic compositions were highly enriched, consistent with microbial SO4(2-) reduction under hydrologic isolation. An analogous microbial couple and similar abiogenic gas chemistry have been reported recently for hydrothermal carbonate vents of the Lost City near the Mid-Atlantic Ridge (D. S. Kelly et al., Science 307:1428-1434, 2005), suggesting that these features may be common to deep subsurface habitats (continental and marine) bearing this geochemical signature. The geochemical setting and microbial communities described here are notably different from microbial ecosystems reported for shallower continental subsurface environments.