饶阳工区微生物控制油井结蜡技术及其现场应用

简述了微生物清防蜡原理及菌种筛选。所用菌种为已商品化的4种兼性厌氧的烃氧化菌，生存温度0－100℃，生长温度20－60℃，最佳生长温度30－45℃，40℃时在饶阳3口井的原油中培养40h，菌数达到峰值，培养80h后菌数仍保持高值。将一种实验菌与3口井的原油在35－40℃培养48h后，原油45℃粘度降低20％－30％，凝固点降低2－3℃，动、静态防蜡率达93％－97％。介绍了微生物清防蜡的选井原则、工艺程序和方法。10口试验井停止热洗和化学清防蜡，每隔30d通过油套环空将50－150kg菌液稀释后注入井筒泵下，油井功图正常，抽油机电流稳定中有所降低，有5口井产油量略有增加。该技术经济上可行。     

降解原油微生物的筛选及其部分特性的研究

筛选得到4株可利用原油为碳源生长的菌株，初步鉴定为假单胞菌。这4株菌对原油有很好的降黏作用，它们能利用长链烷烃生长，并且能耐受一定的温度、压力和矿化度。其中的M－3菌除降黏外还能产生酸和生物表面活性剂，在以原油为碳源的培养基中进行培养时，可以使培养液的表面张力从72mN/m降至36mN/m，pH值从7.0降至约5.5。实验证明M－3菌液在室内条件下能提高原油采收率8.8%左右，在微生物采油中有很好的应用潜力。图1表4参19（李清心摘）     

采油菌B36的室内研究

从胜利油田油水样中分离出一珠细菌B36 ,该菌耐温耐盐性良好 ,能以烃类作为唯一的碳源。该菌 5 0℃时在含有混合石蜡 (>C16原油馏分 )及其他营养物的培养基中摇床培养 12h后 ,培养液中菌数高达 7× 10 5个 /mL ,培养液表面张力由 6 1mN/m降至 32mN/m ,pH值由 7.0降至 5 .4。由培养 36h的上述培养液中分离出黄色粉末状表面活性物质 ,其产量为 0 .6 5 g/L ,经鉴定为脂肽。该菌分别与原油和混合石蜡在不含混合石蜡的培养液中培养 6d后 ,含水原油 5 0℃粘度由 6 5 .6mPa·s降至 15 .3mPa·s,6 0℃界面张力由 30 .6mN/m降至 1.2mN/m ,无水原油凝固点由 4 4℃降至 36℃ ,初馏点下降 ,终馏点升高 ,可馏出物 (轻组分 )由 4 5 .1%增至 5 7.8% ;混合石蜡 2d降解率为2 .1% ,6d降解率达 2 5 .8%。B36是一种有应用前景的采油微生物     

海洋烃细菌的分离及其特性

采自湄洲湾海域的水样，经富集培养，分别得到256株能在以原油为唯一碳源的平板上生长的细菌。通过培养液的比浊法筛选，获得海鱼弧菌（Vibrio damsela)VD-3菌株。VD-3菌株生长的最佳条件是温度20-30℃和溶液中NaCl的质量分数为3.5%。该菌具有较强的降解原油的能力。采用气相色谱分析法，考察VD-3菌对不同烃类的利用。结果表明，该菌既能降解烷烃，也能降解防烃，对长链烷烃正廿四烷的降解率大于正十六烷。     

石油污染土壤生物修复高效菌的降解特性

 研究了从陕北石油污染土壤中分离的7株菌SY21、SY22、SY23、SY24、SY42、SY43和SY44的生长特性及其对不同烃类的利用能力及除油影响因素。结果表明,受试细菌的生物除油率高,培养13 23h后的活性最高,易于扩大培养;分离菌株均能在石蜡培养基中生长,表明对中长链的烷烃降解能力高,其中菌株SY22、SY23、SY24、SY42和SY43均能利用几种不同的烃类生长,是土壤生物修复的优选菌株。菌株SY22 和SY23最适pH为9.0,菌株SY21 和SY42最适pH为7.0;污染强度为1000 1500mg/L、氮源为硝酸铵时,石油烃的降解率最佳;对具有PAH降解能力的SY22、SY42和SY23菌株而言,添加淀粉和葡萄糖为碳源提高了菌株对原油的降解率。土壤中Fe2+、Mn2+的存在对细菌降解石油烃影响不大,但Ni2+、Co2+金属离子会降低分离菌株的除油能力。     

微生物防蜡技术在双河油田的应用

简述了油井微生物防蜡原理。筛选出了可在无机盐存在下以固体石蜡为碳源生长的清防蜡菌种AD-4。菌液含菌>108个/mL。该菌液与9口油井的沉积蜡在65℃作用7天后,沉积蜡凝固点(27.5~55.0℃)下降1.5~8.5℃,菌液pH值由7.4降至6.2~6.8,表面张力(71.0~76.1 mN/m)下降34.2%~49.3%。一口井的原油在60℃与菌液作用8天后,C25以上组分减少,轻组分增多,表明该菌为烃降解菌。在双河区3口井进行先导性试验,在安棚区8口井和下三门区6口井进行适应性试验,AD-4菌液和培养基从套管加入,确定加剂周期为30天,菌液初次加量为300~450 kg,逐次递减至维持量80~150 kg,井温较高时菌液加量较大。5口井生产数据表明加菌液后抽油泵负荷和电流减小,检泵周期延长。该清防蜡菌适用条件为:矿化度<1×104mg/L,含水30%~90%,油层温度<90℃。图1表3参2。     

微生物清防蜡采油技术在王541地区的应用

王541断块原油含蜡量高(10％～35％)，油井井筒结蜡严重，热洗作业频繁。将胜利采油院提供的F18菌组用于区块油井清防蜡取得了良好效果。F18菌组由BS6、B10-1、A3-1、TH1-1、B9—3、N5等6种菌组成，以石蜡为碳源生长，降解石蜡，产生物表面活性剂，阻止蜡晶生长。在室内实验中，F18使11口井产出的原油粘度降低16．5％～59．0％，凝固点降低2～7℃。在现场试验中菌液注入油井环空。在3口井上进行间隔15天、为期3个月的定期注菌液清防蜡先导试验，效果良好。在正式试验中将区块33口井分为4大类，第一类5口井只注菌液；第二类5口井注菌液并热水洗井，洗井周期60天；第三类22口井注菌液并热水洗井，洗井周期45天；第四类1口井(对比井)不加剂不热洗；注菌液周期20天，第一次注0．3吨，以后每次注0．2吨。注菌液井产出的原油，①凝固点略有下降；②原油族组成无明显变化；③轻质饱和烃增多而重质饱和烃减少；井下作业时发现8口注菌液井中只有1口轻微结蜡，而对比井生产85天后管杆严重结蜡。此清防蜡技术适用于含水20％～80％的含水油井，在含水10％的油井中效果不好。图3表3参1。     

采油复合菌的驱油机理

为改善宋芳屯油田的生产状况,从芳6区筛选到了3株以烃类为唯一碳源、兼性厌氧的驱油微生物菌种,并对其与原油的作用、代谢产物对原油物性的影响以及提高采收率的能力进行了研究。研究表明:这3株菌均具有产表面活性剂、产酸、产气活性,最适生长温度相近;它们均可以烃类为碳源、在油层温度下正常繁殖、兼性厌氧生活。组成的复合菌对原油中饱和烃、非烃等基本上都有降解作用。岩心试验表明,复合菌能显著提高采收率,并能在多孔介质中改善原油物性,使原油黏度、油水界面张力、pH值及蜡含量明显降低。该菌种能明显改善原油物性,具有很强的驱油活性。     

克拉玛依油田微生物驱油室内实验研究

从克拉玛依油田井场筛选出１３株兼性厌氧菌，从中复壮出６株菌，筛选出的菌种在克拉玛依油田油藏环境下能以原油烃类为碳源进行生长和繁殖；可以降低原油粘度和倾点，降低油水界面张力。研究发现，菌种能够代谢原油中的较长链烷烃为短链烷烃，同时能够产酸、产气、产表面活性物质。对筛选出的３种菌进行了室内微生物驱油模拟试验，提高水驱后采收率１０％～１５％（ＯＯＩＰ）。     

微生物清蜡降粘采油技术在垦90断块油田的应用

将由垦 90断块油水样中培养出的 12种厌氧、嗜热、耐压菌种按最突出的性能划分为降粘、降蜡、乳化 (降粘 )等菌种 ,分别作用于该断块 4口井产出的原油 ,一些菌使高蜡高凝高粘原油凝固点下降 4 .5～ 9.0℃ ,5 0℃粘度下降16 %～ 5 2 % ,一些菌使原油含蜡量下降 4 %～ 14 % ,一种菌使原油烃主碳峰由C2 4移至C16。考察了这些菌的生长条件 :水矿化度一般 <1.0× 10 5mg/L ,最高达 1.5× 10 5mg/L ;pH值一般 5～ 9,耐酸菌为 5 .5 ,耐碱菌为 7.5。在垦 90断块油藏 (温度 10 0～ 110℃ )的 4口油井中通过油套环空注入由 3种或 4种菌组成的混合菌液 ,一次 30 0～ 10 0 0kg ,使试验井抽油机负荷减小 ,管线回压降低 ,免修期延长 ,一些井产油量有所增加     

抑制油田生产系统中硫酸盐还原菌的方法

在油田开采、集输和注水系统中,硫酸盐还原菌(Sulfate Reducing Bacteria,SRB)是引起微生物腐蚀的主要因素之一。文章对SRB抑制方法的研究进展状况进行综述,着重讨论作为抑制SRB重要措施的杀菌剂的研究进展和新型杀菌剂的开发方向,展望微生物腐蚀研究的发展趋势。     

油田用Fe(Ⅵ)化合物杀菌剂性能评价试验研究

以油田杀菌剂的研制和开发为背景，以制备环境友好杀菌剂并研究其应用为目的，开发了新型油田杀菌剂，并对其性能进行评价。Fe（Ⅵ）化合物杀菌剂是以高铁酸盐（K2FeO4）为主剂的一种氧化型杀菌剂，对各种细菌具有广泛的杀灭作用，可以杀灭油田水中硫酸盐还原菌、腐生菌、铁细菌等。在室内合成该杀菌剂，然后取K2FeO4固体粉末加2～6 mol／L KOH水溶液，配制成0．1％～1％碱性溶液，即为杀菌剂工作液；最后采用绝迹稀释法，测定加杀菌剂前后水样中硫酸盐还原菌、腐生菌和铁细菌的含量，计算杀菌率，评价杀菌效果，确定了杀菌剂在油田水杀菌过程中的使用条件，并从杀菌的角度初步验证了该杀菌剂对油田水中硫酸盐还原菌、铁细菌、腐生菌的杀菌功效。     

Synthesis,characterization and comparative study of nano-Ag–TiO2 against Gram-positive and Gram-negative bacteria under fluorescent light

Currently, some of the major problems affecting the world are air pollution as well as microbial contamination. Every time we breathe, we are risking our lives by inhaling dangerous chemicals and biological contaminants that have found their way into the air. Therefore this work focuses on the antibacterial activity of Ag–TiO₂ to overcome the microbial contaminant and infectious disease. Ag–TiO₂ nanosolution were synthesized by sol–gel method and found to be an effective visible light driven photocatalyst. The nanosolutions were characterized by X-ray diffraction (XRD), transmittance electron microscopy (TEM), photoluminescence (PL) spectra, and X-ray photoelectron spectroscopy (XPS). At the concentration of 0.2–0.1 M, Ag–TiO₂ caused 100% inhibition of bacterial growth. The antibacterial efficacy of Ag–TiO₂ was evaluated with two kinds of bacteria; Gram-positive and Gram-negative. The colony count of Ag–TiO₂ against Gram-negative were evaluated with stain such as Escherichia coli, Pseudomonas aeruginosa and Shigella while for Gram-positive were investigated with Staphylococcus aureus, Bacillus subtilis, Bacillus cereus. Colony count results indicated that Ag–TiO₂ able to kill bacteria at the lowest concentration of 0.05 M that contains 0.06 mol % Ag. From the SEM and TEM observation, the survival of the Gram-positive was low and the decomposition was rapid as compared to Gram-negative bacteria.     

Bottom-Up法计算油田注入水中三种细菌的不确定度

为保持地下压力,在油田开采过程中需向地下油层中注水,注入水中最为常见的三种细菌是腐生菌、铁细菌和硫酸还原菌,这些细菌的存在会降低注入水水质并腐蚀管道,影响油田开采。为客观评价三种细菌检测的准确度,采用绝迹稀释法检测技术,利用Bottom-Up法,通过21组数据对三种细菌检测方法的不确定度进行计算。结果表明,绝迹稀释法检测油田注水三种细菌的不确定度分别为:硫酸还原菌±4 mL^-1,腐生菌±6 mL^-1,铁细菌±16 mL^-1。通过Bottom-Up法对检测不确定度进行评估,可真实、准确地反应油田注入水中三种细菌的实际含量范围,为油田注水达标提供技术保障。     

油气微生物勘探法

本文叙述了油气微生物勘探(MPOG)的发展历史,系统总结了MPOG的理论基础,包括MPOG的原理、物理-化学基础、微生物学基础及其理论模型,简述了MPOG的方法流程。根据国内外MPOG实践经验,论述了MPOG不受地下复杂地质条件的影响,特别适合于非构造油气藏等的主要特点,详细讨论了MPOG受油气藏性质、地层压力等影响因素,并列举了德国、美国及中国的成功实例,最后论述了MPOG的发展趋势。实践证明,采用国外研究机构—高校—油田三结合的方式是引进国外先进技术的有效途径。同时,重视培养和造就一批地质学—微生物学或微生物学、地质学多学科专门人才,发展中国自己的成套地质生物技术,必将对我国大幅度提高油气勘探成功率产生深远的影响。     

Gemini表面活性剂的杀菌性能

采用双子表面活性剂16—4—16、十二烷基二甲基苄基氯化铵1227、异噻唑啉酮3种杀菌剂杀灭回注水中的硫酸盐还原菌（SRB）、铁细菌（FB）及腐生菌（TGB）3种细菌,并用绝迹稀释法对杀菌前后菌数进行测定。实验结果表明,杀菌剂双子表面活性剂和杀菌剂1227的杀菌效率均达到企业标准,且杀菌剂双子表面活性剂的杀菌效率达到99％以上,具有良好的耐药性,可将杀菌剂1227与杀菌剂双子表面活性剂交替应用于现场。     

Microbial diversity and functionally distinct groups in produced water from the Daqing Oilfield, China

The microbial community structure and functionally distinct groups in three kinds of produced water samples from the shallow,mesothermic and low-salinity Daqing oil reservoir were systematically evaluated using both culture-dependent and culture-independent methods.Sequence analysis of the 16S rRNA genes indicated that the bacterial library was dominated by Acinetobacter and Arcobacter and the archaeal community was dominated by Methanosaeta and Methanolinea.Two isolated methanogens were closely related with Methanothermobacter thermautotrophicus and Methanoculleus receptaculi.The fermentative bacteria were identified as Pseudomonas,Haloanaerobium,Alcalibacter,Arcobacter,and Pannonibacter.The predominant nitrate-reducing bacteria fell within the genus Pseudomonas.The dominant members of the cultured hydrocarbon-oxidizing bacteria were phylogenetically associated with Micrococcus,Pseudomonas,and Bacillus.Enrichments of biosurfactants and biopolymer producing groups mainly yielded Pseudomonas,Bacillus,and Acenitobacter-related members.The functional groups related to polymer degradation were also affiliated with Pseudomonas and Bacillus.Results from this study provide the fresh insight into the diversity of microbial communities in Daqing petroleum reservoirs.The vast pool of functional strains retrieved in this study was presumed to include the promising strains that could be applied in microbial-enhanced oil recovery in future.     

油田注水系统中的细菌类型及抑制方法

详细介绍了油田注水系统中主要存在的细菌类型、生存条件、危害机理等，并总结概括了杀菌剂的种类、选用原则和投加方法，在最后，对油田注水用杀菌剂的发展方向进行了展望。     

Use of a submerged yeast culture and lysozyme for the treatment of bacterial contamination during biological aging of sherry wines

The aim of this study was to determine the effect of a submerged culture of “flor” velum yeast and lysozyme as treatments against bacterial contamination by lactic acid and acetic acid bacteria during the biological aging of sherry wine. Different submerged culture treatments with lysozyme were performed. To assess these treatments, we performed laboratory assays with wine spoiled by lactic acid bacteria (LAB) and/or acetic acid bacteria (AAB) selected from the “solera” sherry wine cellar system. The results indicate that lysozyme should be added to submerged culture conditions after the start of the yeast growth phase. The application of lysozyme at a dose of 12.5 g/hL and submerged culture conditions was very effective for the treatment of advanced heterolactic fermentation during the biological aging of wines, thereby inhibiting the LAB population and reducing the volatile acidity of the wines (<0.1 g/L). Similarly, the submerged culture was a very effective tool for the treatment of AAB contamination of recovering wines with high volatile acidity and the removal of AAB without alcohol correction of the wine. The combined lysozyme and submerged culture treatment was very effective at removing bacteria from the wine, reducing volatile acidity and preventing acetification.     

Effectiveness of several chemical decontamination treatments against Gram-negative bacteria on poultry during storage under different simulated cold chain disruptions

Five chemical compounds – 12% trisodium phosphate (TSP), 1200 ppm acidified sodium chlorite (ASC), 2% citric acid (CA), 220 ppm peroxyacids (PA) and 50 ppm chlorine dioxide (CD) – were analyzed to determine their effectiveness against Gram-negative bacteria (Salmonella enterica serotype Enteritidis, Yersinia enterocolitica, Escherichia coli and Pseudomonas fluorescens) artificially inoculated on skinless chicken legs. Samples were stored for 120 h under three different simulated cold chain disruptions: T1 (12 h at 1 ± 1 °C, followed by 6 h at 15 ± 1 °C and then 102 h at 4 ± 1 °C), T2 (18 h at 1 ± 1 °C, 6 h at 15 ± 1 °C and 96 h at 10 ± 1 °C) or T3 (18 h at 4 ± 1 °C, 6 h at 20 ± 1 °C and 96 h at 7 ± 1 °C). Microbiological analyses and pH determinations were carried out at 0, 24, 72 and 120 h of storage. TSP, ASC and CA significantly (P < 0.05) reduced microbial populations, relative to control (untreated) samples. TSP and ASC caused higher (P < 0.05) reductions of S. enterica than CA under moderate temperature abuse conditions (T2 and T3). TSP and ASC (T2) or TSP and CA (T1 and T3) were the most effective compounds against E. coli. TSP and ASC showed the highest antimicrobial effect (P < 0.05) at mild temperature abuse conditions (T1) for Y. enterocolitica, while ASC was the most active compound against this pathogen under T2 and T3 conditions. TSP caused the largest average reductions of Ps. fluorescens at T1. PA and DC originated scant microbial reductions under all of the conditions proposed. These findings extend current knowledge about these chemical decontaminants, and may help the Regulatory Authorities in their decisions about the poultry decontamination authorization process within the European Union.