Stephen J R, Chang Y J, Gan Y D, Peacock A, Pfiffner S M, Barcelona M J, White D C, Macnaughton S J
Center for Environmental Biotechnology, The University of Tennessee, Knoxville 37932, USA.
Environ Microbiol. 1999 Jun;1(3):231-41. doi: 10.1046/j.1462-2920.1999.00030.x.
The impact of pollution on soil microbial communities and subsequent bioremediation can be measured quantitatively in situ using direct, non-culture-dependent techniques. Such techniques have advantages over culture-based methods, which often account for less than 1% of the extant microbial community. In 1988, a JP-4 fuel spill contaminated the glacio-fluvial aquifer at Wurtsmith Air Force Base, Michigan, USA. In this study, lipid biomarker characterization of the bacterial and eukaryotic communities was combined with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis of the eubacterial community to evaluate correlation between contaminant (JP-4 fuel) concentration and community structure shifts. Vadose, capillary fringe and saturated zone samples were taken from cores within and up- and down-gradient from the contaminant plume. Lipid biomarker analysis indicated that samples from within the plume contained increased biomass, with large proportions of typically gram-negative bacteria. Outside the plume, lipid profiles indicated low-biomass microbial communities compared with those within the initial spill site. 16S rDNA sequences derived from DGGE profiles from within the initial spill site suggested dominance of the eubacterial community by a limited number of phylogenetically diverse organisms. Used in tandem with pollutant quantification, these molecular techniques should facilitate significant improvements over current assessment procedures for the determination of remediation end-points.
利用直接的、不依赖培养的技术，可以在原位定量测量污染对土壤微生物群落的影响以及后续的生物修复情况。这类技术比基于培养的方法更具优势，基于培养的方法所涵盖的微生物群落通常不到现存微生物群落的1%。1988年，一次JP - 4燃油泄漏污染了美国密歇根州伍尔史密斯空军基地的冰川河流含水层。在本研究中，将细菌和真核生物群落的脂质生物标志物特征与真细菌群落的聚合酶链反应 - 变性梯度凝胶电泳（PCR - DGGE）分析相结合，以评估污染物（JP - 4燃油）浓度与群落结构变化之间的相关性。从污染物羽流内部以及羽流上下游的岩芯中采集了渗流带、毛细边缘带和饱和带的样本。脂质生物标志物分析表明，羽流内部的样本生物量增加，其中大部分是典型的革兰氏阴性菌。在羽流外部，脂质谱显示与初始泄漏点内的微生物群落相比，生物量较低。从初始泄漏点内的DGGE图谱中获得的16S rDNA序列表明，真细菌群落由数量有限的、系统发育多样的生物体主导。这些分子技术与污染物定量结合使用，应该会比目前用于确定修复终点的评估程序有显著改进。
