油藏微生物及其在石油工业中的应用

油藏是一种特殊环境,其高温、高压、高矿化度、无氧、多孔介质及其流体等因素对微生物的存活及生长繁殖都会产生明显影响。油藏极端环境对微生物群落结构组成和数量也会有大的影响。该文介绍了与微生物采油关系紧密的微生物群落的生理生化特征及其多样性。主要有烃降解菌、发酵菌、硫酸盐还原菌及产甲烷菌。还介绍了激活本源微生物和筛选外源微生物在提高石油采收率方面的应用。最后就分子生物技术在研究油藏中可培养微生物和未培养微生物中的应用进行了讨论。     

高温油藏内源微生物及其提高采收率潜力研究

大港孔店油田油藏特征、流体和微生物性质分析结果表明, 属于高温生态环境, 地层水矿化度较低, 氮、磷浓度低, 而且缺乏电子受体, 主要的有机物来源是油气。油田采用经过除油处理的油藏产出水回注方式开发, 油层中存在的微生物类型主要是厌氧嗜热菌, 包括发酵菌(102个/mL~105个/mL), 产甲烷菌(103个/mL); 好氧菌主要存在于注水井周围。硫酸盐还原菌(SRB)还原速率0.002 mg S2-/(L·d) ~18.9 mg S2-/(L·d), 产甲烷菌产甲烷速率0.012 mgCH4/(L·d)~16.2 mgCH4/(L·d)。好氧菌能够氧化油形成生物质, 部分氧化产物为挥发性脂肪酸和表面活性剂。产甲烷菌在油氧化菌液体培养基中产生CH4, CO2为好氧微生物和厌氧微生物的共同代谢产物。这些产物具有提高原油流动性的作用。用示踪剂研究了注入水渗流方向。通过综合分析, 油藏微生物具有较大的潜力, 基于激活油层菌的提高采收率方法在该油田是可行的。     

高温油藏内源微生物及其提高采收率潜力研究

大港孔店油田油藏特征、流体和微生物性质分析结果表明,属于高温生态环境,地层水矿化度较低,氮、磷浓度低,而且缺乏电子受体,主要的有机物来源是油气.油田采用经过除油处理的油藏产出水回注方式开发,油层中存在的微生物类型主要是厌氧嗜热菌,包括发酵菌(102个/mL～105个/mL),产甲烷菌(103个/mL);好氧菌主要存在于注水井周围.硫酸盐还原菌(SRB)还原速率0.002 μg S2-/(L·d)～18.9 μg S2-/(L·d),产甲烷菌产甲烷速率0.012 μgCH4/(L·d)～16.2 μgCH4/(L·d).好氧菌能够氧化油形成生物质,部分氧化产物为挥发性脂肪酸和表面活性荆.产甲烷菌在油氧化菌液体培养基中产生CH4,CO2为好氧微生物和厌氧微生物的共同代谢产物.这些产物具有提高原油流动性的作用.用示踪剂研究了注入水渗流方向.通过综合分析,油藏微生物具有较大的潜力,基于激活油层茵的提高采收率方法在该油田是可行的.     

采油微生物在微驱过程中的生长、运移及分布规律

[目的]深入了解现场微生物驱油机理、效果评价标准及影响因素.[方法]结合现场微生物驱油过程产出液的跟踪监测及室内物模实验对微生物在地层中的生长繁殖、运移及分布规律进行研究.[结果]结果表明,通过从水井注入的外源微生物在油藏中能够有效生长繁殖,而且注入的营养液也能够激活内源微生物,但由于地层渗透率及营养液浓度的影响,产出液菌浓要比注入菌浓低1-2个数量级;葡萄糖的快速降解以及地层对微生物的过滤及吸附作用使大量的微生物停留在近井地带,仅有部分微生物能够从生产井采出,而且其运移速度要比营养液慢.[结论]地层渗透率和产出液中营养物浓度是影响微生物数量及分布的两个关键因素,现场微生物驱油产出液中的菌浓一般很难达到106个/mL以上,该研究结果对微生物驱油技术的发展和应用具有重要意义.     

油藏极端环境中的微生物

油藏属于高温、高压、高矿化度以及油、气、水共存的极端环境,与之相适应,油藏中孕育着丰富多样的微生物.介绍了油藏极端环境、油藏微生物的一般特征、典型的微生物生理群以及随油田开发微生物分布的变化.     

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。     

不同激活剂对油藏内源微生物群落结构的影响

目的筛选大庆油田聚驱后油藏内源微生物采油激活剂的类型。方法在动态物理模拟实验模型基础上,利用基于16S rDNA的基因克隆文库方法,分析了两组激活剂配方激活的微生物群落结构。结果配方1激活的微生物菌群主要有假单胞菌属(75%)、深海弯曲菌属(18%)、陶厄菌属(6%);配方2激活的微生物菌群主要有陶厄菌属(50%)、假单胞菌属(26%)、螺杆菌属(15%)、梭菌属(9%)。结论激活剂配方2更适合大庆油田聚驱后油藏内源微生物采油。     

大庆聚驱后油藏微生物群落结构分析

目的:进一步研究大庆聚驱后油藏微生物种类.方法:应用T-RFLP方法比较系统地分析比较了聚驱后油藏微生物多样性,分析了与采油功能相关的主要微生物组成,初步研究了聚驱后油藏微生物分布特点及规律.结果:利用聚驱后油藏细菌群落16S rDNA构建克隆文库,共挑取了49个阳性克隆,T-RFLP分型结果,共出现21个不同类型.所得的序列在Genbank中比对的结果表明细菌克隆可分为5个类群,分属于变形杆菌、拟杆菌、脱铁杆菌、厚壁菌以及未确定分类的类群;同时构建了聚驱后油藏古菌群落16S rDNA的克隆文库,共挑取了48个阳性克隆,根据T-RF分型结果共有5个类群,构建的古菌克隆文库的12个阳性克隆分属5个不同的类群,这些克隆和产甲烷菌亲缘关系很近.结论:该研究为大庆油田本源微生物采油技术的应用提供了可靠的理论依据.     

中高温油藏内源微生物厌氧激活

为了明确中高温油藏内源微生物厌氧激活产气的特点,在胜利油田选取了11个区块开展产气研究,温度范围分别为55~65℃、65~79℃、79~95℃。在模拟油藏条件下厌氧激活发现,低于79℃时,油藏内源微生物普遍能被激活并代谢产气;而高于79℃时,无明显甲烷气产生。利用不同碳源激活后发现,H_2/CO_2为碳源时,最大产甲烷速率可达1 500μmol/(g·d),显著高于乙酸钠和淀粉,这表明中高温油藏内产甲烷古菌以氢营养型为主。进一步对正理庄正南区块内源微生物群落组成研究,发现厌氧激活后古菌中Methanobacterium被激活成为优势菌群,有利于在油藏内代谢产甲烷气。厌氧激活前油水样中优势菌都是Pseudomonas,不同激活剂条件下优势菌群发生明显变化,H2/CO2和淀粉以激活Thermotoga为主,而乙酸钠则主要激活了油井中的Deferribacter。此外,该区块不同油井在相同激活条件下,其优势菌群趋向于一致,但油井和水井中优势菌群存在明显差异。通过对中高温油藏内源微生物厌氧激活的研究,为进一步提高微生物采油的效果奠定了基础。     

厌氧产表面活性剂微生物提高原油采收率的研究进展

微生物强化采油(microbial enhanced oil recovery,MEOR)是近年来在国内外发展迅速的一项提高原油采收率技术。微生物在油藏中高效生产表面活性剂等驱油物质是微生物采油技术成功实施的关键之一。然而,油藏的缺/厌氧环境严重影响好氧表面活性剂产生菌在油藏原位的生存与代谢活性;油藏注空气会增加开采成本,且注入空气的作用时效和范围难以确定。因此,开发厌氧产表面活性剂菌种资源并强化其驱油效率对于提高原油采收率具有重要意义。本文综述了国内外近年来利用厌氧产表面活性剂微生物提高原油采收率的研究进展,简述了微生物厌氧产表面活性剂的相关驱油机理、菌种资源开发现状以及油藏原位驱油应用进展,并对当前的研究提出了一些思考。     

高糖高胰岛素对去甲肾上腺素诱导的心肌细胞肥大的影响

目的:利用体外培养的乳鼠心肌细胞,观测高浓度胰岛素和高浓度葡萄糖作用下对去甲肾上腺素诱导的心肌细胞肥大的影响,并探讨其可能作用机制.方法:以培养的乳鼠心肌细胞为模型分组给药后,用显微镜目镜计数心肌细胞搏动的频率;用Lowrys法测心肌细胞的蛋白质含量;用消化分离法,利用计算机图象分析系统测心肌细胞的体积;用[3H]leucine标记法测定心肌细胞蛋白的合成.结果:与对照组相比,去甲肾上腺素(NE)组、高糖组、高胰岛素组心肌细胞蛋白含量、体积、蛋白合成均有明显增加,而与高糖加NE组和高糖高胰岛素组相比较,高糖高胰岛素加NE组心肌细胞蛋白含量、体积、蛋白合成增加则更为显著.结论:单纯高浓度胰岛素培养可促进心肌细胞肥大,同时用高糖高胰岛素联合培养,可使去甲肾上腺素诱导的心肌细胞肥大作用进一步增强.     

Diet-Induced Changes in AChE Activity after Long-Term Exposure

In the present study we investigated a potential mechanism by which high sugar (HS) and high fat (HF) diets could affect acetylcholinesterase (AChE) activity. The treatment with HS and HF diet was done for six months on male and female rats. The results showed decreased hippocampal AChE activity in male and females receiving HS and HF diets (HS 24% and 36%; HF 38% and 32%, males and females, respectively; P < 0.05). The activity in the cerebral cortex was reduced in males (49 and 40%) and females (19 and 17%) (P < 0.05) on HS and HF diets, respectively. In the hypothalamus AChE activity was decreased on HS diet in males (46%) and female (25%) (P < 0.05) and also on HF diet in males (34%) and females (21%) (P < 0.05). However, in the cerebellum no changes in AChE activity were observed. These results indicate that HS and HF diets produced mainly inhibition in acetylcholine degradation. It probably indicates a chronic alteration induced by these diets on the cholinergic system.     

Effects of instantaneous and growth CO2 levels and abscisic acid on stomatal and mesophyll conductances

C₃ photosynthesis is often limited by CO₂ diffusivity or stomatal (g_s) and mesophyll (g_m) conductances. To characterize effects of stomatal closure induced by either high CO₂ or abscisic acid (ABA) application on g_m, we examined g_s and g_m in the wild type (Col‐0) and ost1 and slac1‐2 mutants of Arabidopsis thaliana grown at 390 or 780 μmol mol^?1 CO₂. Stomata of these mutants were reported to be insensitive to both high CO₂ and ABA. When the ambient CO₂ increased instantaneously, g_m decreased in all these plants, whereas g_s in ost1 and slac1‐2 was unchanged. Therefore, the decrease in g_m in response to high CO₂ occurred irrespective of the responses of g_s. g_m was mainly determined by the instantaneous CO₂ concentration during the measurement and not markedly by the CO₂ concentration during the growth. Exogenous application of ABA to Col‐0 caused the decrease in the intercellular CO₂ concentration (C_i). With the decrease in C_i, g_m did not increase but decreased, indicating that the response of g_m to CO₂ and that to ABA are differently regulated and that ABA content in the leaves plays an important role in the regulation of g_m.     

Formation of high density lipoproteins containing both apolipoprotein A-I and A-II in the rabbit

Human plasma HDLs are classified on the basis of apolipoprotein composition into those that contain apolipoprotein A-I (apoA-I) without apoA-II [(A-I)HDL] and those containing apoA-I and apoA-II [(A-I/A-II)HDL]. ApoA-I enters the plasma as a component of discoidal particles, which are remodeled into spherical (A-I)HDL by LCAT. ApoA-II is secreted into the plasma either in the lipid-free form or as a component of discoidal high density lipoproteins containing apoA-II without apoA-I [(A-II)HDL]. As discoidal (A-II)HDL are poor substrates for LCAT, they are not converted into spherical (A-II)HDL. This study investigates the fate of apoA-II when it enters the plasma. Lipid-free apoA-II and apoA-II-containing discoidal reconstituted HDL [(A-II)rHDL] were injected intravenously into New Zealand White rabbits, a species that is deficient in apoA-II. In both cases, the apoA-II was rapidly and quantitatively incorporated into spherical (A-I)HDL to form spherical (A-I/A-II)HDL. These particles were comparable in size and composition to the (A-I/A-II)HDL in human plasma. Injection of lipid-free apoA-II and discoidal (A-II)rHDL was also accompanied by triglyceride enrichment of the endogenous (A-I)HDL and VLDL as well as the newly formed (A-I/A-II)HDL. We conclude that, irrespective of the form in which apoA-II enters the plasma, it is rapidly incorporated into spherical HDLs that also contain apoA-I to form (A-I/A-II)HDL.     

银耳子实体多糖高温高压提取工艺研究

通过对银耳子实体多糖高温高压提取工艺中,提取温度、料液比、提取时间,提取次数等影响因素的实验分析,确定银耳子实体多糖高温高压提取的最佳条件为:提取温度110℃(对应压力0.04 MPa);料液比为1∶70;提取时间为2 h;提取次数为2次,在此条件下,银耳子实体多糖提取率可达36.5%。     

Apolipoprotein M expression increases the size of nascent pre�� HDL formed by ATP binding cassette transporter A1

Apolipoprotein M (apoM) is a novel apolipoprotein that is reportedly necessary for preβ HDL formation; however, its detailed function remains unknown. We investigated the biogenesis and properties of apoM and its effects on the initial steps of nascent preβ HDL assembly by ABCA1 in HEK293 cells. Transiently transfected apoM was localized primarily in the endomembrane compartment. Pulse-chase analyses demonstrated that apoM is inefficiently secreted, relative to human serum albumin, and that ∼50% remains membrane-associated after extraction with sodium carbonate, pH 11.5. To investigate the role of apoM in nascent preβ HDL formation, ABCA1-expressing or control cells, transfected with empty vector, apoM, or C-terminal epitope-tagged apoM (apoM-C-FLAG), were incubated with ¹²⁵I-apoA-I for 24 h. Conditioned media were harvested and fractionated by fast-protein liquid chromatography (FPLC) to monitor HDL particle size. Preβ HDL particles were formed effectively in the absence of apoM expression; however, increased apoM expression stimulated the formation of larger-sized nascent preβ HDLs. Immunoprecipitation with anti-apoA-I antibody followed by apoM Western blot analysis revealed that little secreted apoM was physically associated with preβ HDL. Our results suggest that apoM is an atypical secretory protein that is not necessary for ABCA1-dependent preβ HDL formation but does stimulate the formation of larger-sized preβ HDL. We propose that apoM may function catalytically at an intracellular site to transfer lipid onto preβ HDL during or after their formation by ABCA1.     

康脂口服液药效学研究

通过体内外血栓形成实验证明康脂口服液可明显延长大鼠血栓形成时间,减少血栓长度和重量.对高血脂大、小鼠血总脂、总胆固醇、甘油三酯均有不同程度降低作用,对高血脂大鼠低密度脂蛋白增高有明显抑制作用.此外可增加麻醉犬脑血流量,降低脑血管阻力,减轻脑缺血后脑水肿,降低血粘度,改善微循环,降低血小板聚集率.     

气温和根区温度对葡萄叶片光合荧光特性的影响

为探究气温和根区温度对葡萄(Vitis vinifera)叶片光合荧光特性的影响, 以一年生巨峰葡萄为试材, 设置对照、高气温、高根区温度和两者交叉作用共4组处理。结果表明, 相较于对照和高气温, 高根区温度以及交叉处理叶片最大光化学效率(F_v/F_m)降低更明显; 与对照相比, 高根区温度以及高气温与高根区温度交叉处理下光系统II (PSII)实际光化学效率Y(II)显著降低, 非调节能量耗散的量子产量Y(NPQ)及Q_A氧化还原状态(1-q_P)值显著上升。同时, 高根区温度以及高气温与高根区温度交叉处理显著增加了J点的可变荧光(V_j), 而用于电子传递的量子产额(φ_Eo)及性能指数(PI_ABS)显著降低。此外, 高根区温度以及高气温与高根区温度交叉处理下单位面积有活性的反应中心数目(RC/CS_m)也显著下降, K点相对可变荧光(W_k)明显上升。综上所述, 高根区温度是高气温与根区高温交叉胁迫的主导因子, PSII受体侧是主要的伤害位点, 高气温加剧了高根区温度对PSII造成的伤害。     

EXTREMELY HIGH GROWTH RATE OF THE SMALL DIATOM CHAETOCEROS SALSUGINEUM ISOLATED FROM AN ESTUARY IN THE EASTERN SETO INLAND SEA,JAPAN1

Small single‐celled Chaetoceros sp. are often widely distributed, but frequently overlooked. An estuarine diatom with an extremely high growth potential under optimal conditions was isolated from the Shinkawa‐Kasugagawa estuary in the eastern part of the Seto Inland Sea, western Japan. It was identified as Chaetoceros salsugineum based on morphological observations. This strain had a specific growth rate of 0.54 h^?1 at 30°C under 700 μmol · m^?2 · s^?1 (about 30% of natural maximal summer light) with a 14:10 L:D cycle; there was little growth in the dark. However, under continuous light it grew at only 0.35 h^?1 or a daily specific growth rate of 8.4 d^?1. In addition, cell density, chlorophyll a, and particulate organic carbon concentrations increased by about 1000 times in 24 h at 30°C under 700 μmol · m^?2 · s^?1 with a 14:10 L:D cycle, showing a growth rate of close to 7 d^?1. This very rapid growth rate may be the result of adaptation to this estuarine environment with high light and temperature. Thus, C. salsugineum can be an important primary producer in this estuary in summer and also an important organism for further physiological and genetic research.