Metagenomics Microbial Characterization of Production and Process Fluids in the Powder River Basin: Identification and Sources of Problematic Microorganisms Associated with SWD Facilities

M. Enzien, Sadie Starustka, Michael J Gurecki, Trinity Fincher-Miller, Bryce Kuhn, Carly Sowecke, Kody B Jones, Kevin O'Sullivan, Keith C. Norris, J. Stidham
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引用次数: 1

Abstract

Inconsistent bacterial control and monitoring led to variability in Salt Water Disposal (SWD) well performance and injectivity creating excess costs in biocide applications and remedial work. A metagenomics study using Whole Genome Sequencing (WGS) was conducted to determine the source(s) of problematic microorganisms throughout the process life cycle: Freshwater> Drilling> Completion> Flowback> Produced water> SWD. A total of 30 metagenomes were collected from the 6 process stages and identification and quantification of the major microbial taxa from each of these stages were identified. "Taxonomy to Function" associations were identified for all the major taxa found in the SWD fluids. WGS was performed on positive Sulfate Reducing Bacteria (SRB) and Acid Producing Bacteria (APB) media bottles inoculated in the field for a Flowback sample. Four of the six major taxa found in SWD samples are considered groups of microorganisms known to cause microbiologically influenced corrosion (MIC): Clostridia, methanogens, SRB and Iron Reducing bacteria. Thermovirga and Thermotagae, were the two most abundant taxa found in SWD samples, both thermophilic halophilic fermenting bacteria. The Fe reducing bacteria Shewanella was only detected in Drilling and SWD fluids suggesting its source was Drilling fluids. Completion fluid metagenome profiles from two separate sites followed similar patterns. During middle of completions Proteobacteria phyla were dominant taxa represented mostly by Pseudomonas. Other abundant phyla were all characteristic of polymer degrading bacteria. None of these taxa were dominant populations identified in SWD waters. Fresh water only shared similar taxa with Drilling and Completion fluids. A few minor taxa from Drilling and Completion stages show up as significant taxa in SWD fluids. The majority of taxa found in SWD samples appear to originate from Flowback and Produced waters, although at lower abundances than found in SWD samples. It cannot be determined if the microorganisms found in Flowback and Produced waters were endemic to the formation or come from contaminated source waters, i.e. process equipment used to store and transport water sources. Petrotoga mobilis was the dominant population of bacteria that grew in both media bottles, 96% and 77% for SRB and APB, respectively, while Petrotoga was detected at 14% in the field sample. The most abundant bacteria detected in field sample were Clostridia (38%) while only 2.7% were detected in APB media. SRB media bottle had 0.18% SRB detected by WGS; APB media had 9% SRB population abundance. No SRB were detected in corresponding field sample or below detectable limits (BDL) for WGS methods (<0.01%). WGS was forensically used to successfully identify type and source of problematic microorganism in SWD facilities. Results from media bottle and field sample comparisons stress the importance of developing improved field monitoring techniques that more accurately detect the dominant microorganisms.
粉末河流域生产和工艺流体的宏基因组学微生物特征:与SWD设施相关的问题微生物的鉴定和来源
不一致的细菌控制和监测导致了盐水处理(SWD)井的性能和注入能力的变化,造成了杀菌剂应用和补救工作的额外成本。利用全基因组测序(WGS)进行了一项元基因组研究,以确定整个过程生命周期中问题微生物的来源:淡水>钻井>完井>返排>采出水> SWD。共收集了6个过程阶段的30个宏基因组,并对每个过程阶段的主要微生物类群进行了鉴定和定量。在SWD液体中发现的所有主要分类群都确定了“分类功能”关联。对现场接种的硫酸盐还原菌(SRB)和产酸菌(APB)阳性培养基瓶进行WGS,获得返排样品。在SWD样品中发现的六个主要分类群中,有四个被认为是已知引起微生物影响腐蚀(MIC)的微生物群:梭状芽孢杆菌、产甲烷菌、SRB和铁还原菌。在SWD样品中发现的两个最丰富的分类群是Thermovirga和Thermotagae,它们都是嗜热嗜盐发酵细菌。铁还原菌希瓦氏菌仅在钻井液和SWD液中检测到,表明其来源为钻井液。来自两个不同地点的完井液宏基因组剖面具有相似的模式。完成中期以变形菌门为优势类群,以假单胞菌为主。其他丰富的门都是聚合物降解菌的特征。这些类群均不是在SWD水域中发现的优势种群。淡水仅与钻井完井液具有相似的分类群。钻井和完井阶段的一些小类群在SWD流体中表现出重要的类群。在社会福利署样本中发现的大部分分类群似乎来自返排和采出水,尽管其丰度低于社会福利署样本。无法确定在返排和产出水中发现的微生物是地层特有的,还是来自受污染的水源,即用于储存和运输水源的工艺设备。在SRB和APB培养基中,Petrotoga mobilis是优势菌群,分别为96%和77%,而在野外样品中,Petrotoga的检出率为14%。野外样品中检出最多的细菌为梭状芽胞杆菌(38%),而APB培养基中检出的细菌仅为2.7%。WGS检测SRB培养基瓶SRB含量为0.18%;APB培养基中SRB种群丰度为9%。在相应的野外样品中未检出SRB或低于WGS方法的可检出限(BDL)(<0.01%)。WGS在法医鉴定中成功鉴定了社署设施中问题微生物的类型和来源。培养基瓶和现场样品比较的结果强调了开发改进的现场监测技术以更准确地检测优势微生物的重要性。
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