ENRICHMENT AND STABILITY OF ENDOGENOUS MICROBIAL COMMUNITY IN CRUDE OIL PHASE OF RESERVOIR-PRODUCED FLUID
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摘要: 油藏采出液的原油相近年来被认为是获得原油降解菌的理想环境,但缺乏对其微生物分离富集的研究。以华北油田6口油井采出液为研究对象,比较了采出液油水两相的微生物群落结构,随后富集了各原油相的内源菌群,同时跟踪了富集过程中的群落演替,并考察了富集菌群对环境因子多次连续扰动的响应。结果表明:原油相的微生物多样性及丰富度均高于水相样品,且两相间的群落结构存在显著差异。P1号、92号和99号油井的原油相中分别含有111、23和9个独特的OTUs。连续富集10代后,富集群落15号、92号和P1号的原油降解率逐步增加并趋于稳定,最高降解率分别为81.9%、71.5%和63.6%。15号群落的优势菌主要包括Brevibacillus (89.3%)、Novibacillus (8.0%) 和Bacillus (1.5%),而92号和P1号富集群落以Brevibacillus为绝对优势菌,其相对丰度高达99%以上。在温度及氧气的多次连续扰动下,P1号群落的微生物组成及丰度均发生变化,优势菌Brevibacillus被抑制,同时Paenibacillus和Aneurinibacillus的相对丰度上升,造成部分中长链烷烃(C18—C29)的积累;但15号和92号群落可维持稳定的烷烃降解能力。上述结果表明,采出液的原油相可以作为原油降解菌的潜在微生物源。Abstract: The crude oil phase of reservoir-produced fluid has been considered as an ideal environment for obtaining crude oil-degrading bacteria in recent years, but there is little research on microbial isolation from the crude oil phase. In this study, the microbial community structure in the aqueous-oil phase of the produced fluid from six oil wells located in the North China Oilfield was compared. Then, the endogenous community in the six crude oil phases was enriched, respectively. At the same time, the community succession in the process of enrichment was dynamically tracked. Finally, the response of the enriched communities to continuous disturbance of environmental factors was investigated. The results showed that the microbial diversity and richness in the crude oil phase were higher than those in the aqueous phase, and there were significant differences in the community structure between the two phases. The crude oil phases from well P1#, 92#, and 99# contained 111, 23, and 9 unique OTUs, respectively. After 10 generations of continuous enrichment, the crude oil degradation rates of enriched communities 15#, 92#, and P1# gradually increased and tended to be stable, and the maximum degradation rate was 81.9%, 71.5%, and 63.6%, respectively. The dominant bacteria in the community 15# mainly included Brevibacillus (89.3%), Novibacillus (8.0%), and Bacillus (1.5%), while Brevibacillus was the absolute dominant bacteria (with a relative abundance of 99%) in the community 92# and P1#. Under the multiple disturbances of temperature and oxygen, the microbial composition and abundance of community P1# changed. The dominant Brevibacillus was suppressed while the relative abundance of Paenibacillus and Aneurinibacillus increased, resulting in the accumulation of medium and long chain alkanes (C18 to C29). However, community 15# and 92# can maintain stable ability of alkane degradation. These results indicated that the crude oil phase of the produced liquid can act as a potential microbial source for exploring crude oil-degrading bacteria.
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Key words:
- crude oil phase /
- microbial community /
- enrichment /
- crude oil degradation /
- environmental disturbance
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