Estimating the Abundance of Endospores of Sulfate-Reducing Bacteria in Environmental Samples by Inducing Germination and Exponential Growth
Estimating the Abundance of Endospores of Sulfate-Reducing Bacteria in Environmental Samples by Inducing Germination and Exponential Growth
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DOI：
10.1080/01490451.2016.1190805
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发表时间：
2017-01-01
影响因子：
2.3
通讯作者：
Jorgensen, Bo Barker
中科院分区：
文献类型：
--
作者：
de Rezende, Julia Rosa;Hubert, Casey R. J.;Jorgensen, Bo Barker
定量区分环境样本中活跃和休眠的微生物种群是一个挑战。在这里，我们提出了一种方法来估计休眠的硫酸盐还原菌（SRB）的丰度，目前在环境样品中的活内生孢子。这是通过诱导内生孢子发芽和指数生长来实现的。我们证明了这种方法在奥胡斯湾，丹麦的温带沉积物中的嗜热SRB。该方法基于测量巴氏灭菌沉积物中的整体硫酸盐还原率（SRR），并基于指数生长纯培养物中已知的硫酸盐还原菌生长产量和细胞大小的平均值计算相关的细胞特异性SRR。提出的方法是一个更快的生物测定比最可能的数量计数，并有可能区分缓慢和快速增长的SRB群体在同一个样品。考虑到内生孢子对细胞透化和裂解所带来的挑战，这种生物测定是有吸引力的，这是定量显微镜和基于核酸提取的策略的先决条件。这些分子方法还依赖于设计靶合适的寡核苷酸探针，而本文提出的方法更广泛地考虑了感兴趣的性状。因此，这一战略提出了一个有益的补充，其他方法在生态调查的微生物群落学和具体的工业应用，如储层酸化和腐蚀风险评估的石油和天然气部门。
It is a challenge to quantitatively distinguish active from dormant microbial populations in environmental samples. Here we present an approach for estimating the abundance of dormant sulfate-reducing bacteria (SRB), present as viable endospores in environmental samples. This is achieved by inducing endospores to germinate and grow exponentially. We demonstrate this approach for thermophilic SRB in temperate sediment from Aarhus Bay, Denmark. The approach is based on measuring bulk sulfate reduction rates (SRRs) in pasteurized sediment and calculating associated cell-specific SRRs based on average values for SRB growth yield and cell size known from exponentially growing pure cultures. The method presented is a faster bioassay than most probable number enumerations and has the potential to distinguish between slow- and fast-growing SRB populations in the same sample. This bioassay is attractive given the challenges posed by endospores with respect to cell permeabilization and lysis, which are prerequisite in quantitative microscopy- and nucleic acid extraction-based strategies. These molecular approaches additionally rely on designing target-appropriate oligonucleotide probes, whereas the method presented here considers the trait of interest more broadly. This strategy thus presents a useful complement to other methods in ecological investigations of microbial biogeography and for specific industrial applications such as reservoir souring and corrosion risk assessments in the oil and gas sector.