Comparing the response of the indigenous microbial community to crude oil amendment in oxic versus hypoxic conditions

Z. G. Griffiths, Andrew D. Putt, J. I. Miller, M. F. Campa, D. Joyner, O. Pelz, Nargiz Garajayeva, M. Ceccopieri, P. Gardinali, Terry C. Hazen
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Abstract

The Caspian Sea is the world’s largest landlocked saline lake which lies between Europe and Asia. This region is particularly known for its large-scale oil reserves, pipelines, and drilling activities, which have contributed to the environmental decline of this lake. In addition to pollution from the petroleum industry, drainage from various river basins brings an influx of residential, industrial, and agricultural effluents that induce eutrophication and hypoxic conditions in deeper, colder waters, creating an oxygen gradient. The temperature and oxygen stratification in this environment has presented a unique opportunity to investigate the potential of the biodegradative processes carried out by the indigenous microbial community. We believe these indigenous microbes possess different metabolic capabilities to degrade oil as they adapted to declining oxygen concentrations and temperatures with increasing depths over a prolonged period. Hence, community structure and composition will vary with depth.Microcosms were set up to observe the indigenous microbial reaction after a 60 ppm native crude oil amendment over 115 days. Surface water microcosms were incubated at 28ºC and aerated while deep water microcosms were incubated at 8ºC under anaerobic conditions. These two environmental conditions represent the temperature and oxygen extremes along the gradient and were selected as we try to simulate the indigenous community’s response to this oil contamination. DNA was extracted and amplified from these microcosms and sequenced. Bioinformatic analysis was performed to track changes in the abundance of taxa present and biodiversity over different time points to show the progression of community structure.All microcosms showed the presence of hydrocarbon-degrading phyla, whose presence is consistent with other reports from oil-enriched environments. However, distinct communities were observed in oxic versus hypoxic microcosms.Orders of Bacteria related to sulfate and nitrogen cycling were found in hypoxic microcosms, indicating a possible mechanism for the anaerobic biodegradation of crude oil. GC-MS analysis of initial and final microcosms also provided evidence of degradation of hydrocarbon fractions in both warm, oxic and cold, hypoxic conditions.
比较本地微生物群落在缺氧和缺氧条件下对原油添加剂的反应
里海是世界上最大的内陆咸水湖,位于欧洲和亚洲之间。该地区尤其以其大规模的石油储备、管道和钻探活动而闻名,这些活动导致了该湖泊环境的恶化。除了石油工业造成的污染外,各流域的排水也带来了大量的居民、工业和农业污水,这些污水在较深、较冷的水域造成富营养化和缺氧状况,形成了氧气梯度。这种环境中的温度和氧气分层为研究本地微生物群落进行生物降解过程的潜力提供了一个独特的机会。我们相信,这些本地微生物拥有不同的新陈代谢能力来降解石油,因为它们适应了随着深度增加而长期下降的氧气浓度和温度。因此,群落结构和组成会随着深度的变化而变化。我们建立了微生态系统,以观察 60 ppm 原生原油添加剂 115 天后本地微生物的反应。表层水微型培养箱在 28ºC 温度下充气培养,而深层水微型培养箱则在厌氧条件下于 8ºC 温度下培养。这两种环境条件代表了沿梯度的极端温度和氧气条件,我们选择这两种环境条件是为了模拟土著群落对石油污染的反应。从这些微生态系统中提取和扩增 DNA 并进行测序。我们进行了生物信息学分析,以跟踪不同时间点上存在的类群丰度和生物多样性的变化,从而显示群落结构的进展。不过,在缺氧和缺氧微生态系统中观察到了不同的群落。在缺氧微生态系统中发现了与硫酸盐和氮循环相关的细菌,这表明原油厌氧生物降解的可能机制。对初始和最终微生态系统进行的气相色谱-质谱分析也提供了碳氢化合物馏分在温暖缺氧和寒冷缺氧条件下降解的证据。
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