Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms
Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms
复制标题
追踪洋葱伯克霍尔德菌 G4 5223-PR1 在含水层微观世界中的反应
DOI：
10.1128/aem.61.2.448-455.1995
复制
发表时间：
1995
影响因子：
4.4
通讯作者：
Andrichard A. Snyder
中科院分区：
文献类型：
--
作者：
K. N. Timmis;Andrichard A. Snyder
为了生物修复(生物增强)目的而将细菌引入环境，需要分析和监测微生物种群动态，以从功效和风险评估的角度确定持久性和活性。洋葱伯克霍尔德氏菌G4 5223-Pr1是一个Tn5插入突变体，它结构性地表达降解三氯乙烯(TCE)的甲苯邻位单加氧酶。G4 5223-PR1在不诱导芳香族化合物的情况下降解三氯乙烯的这种能力可能对含三氯乙烯的含水层和地下水的生物修复有用。因此，模拟含水层沉积物系统和地下水微宇宙被用来监测G4 5223-PR1的存活率。通过间接免疫荧光显微镜、菌落印迹试验和选择培养基上的生长监测G4 5223-PR1在沉积物中的去向。G4 5223-PR1是用一种高度特异的抗该菌脂多糖O-特异性多糖链的单抗进行免疫检测的。G4 5223-PR1在灭菌地下水中存活良好，但在非灭菌地下水微宇宙中观察到G4 5223-PR1细胞数量迅速减少。接种后10d，选择性平板培养和免疫荧光检测均未检测到G45223-PR1细胞。G4 5223-PR1在非灭菌含水层沉积物微宇宙中的存活率更高，尽管经过22天的洗脱后，G4 5223-PR1细胞的数量很少。我们的结果证明了单抗跟踪方法的实用性和生物相互作用在确定引入微生物的持久性方面的重要性。
The introduction of bacteria into the environment for bioremediation purposes (bioaugmentation) requires analysis and monitoring of microbial population dynamics to define persistence and activity from both efficacy and risk assessment perspectives. Burkholderia cepacia G4 5223-PR1 is a Tn5 insertion mutant which constitutively expresses a toluene ortho-monooxygenase that degrades trichloroethylene (TCE). This ability of G4 5223-PR1 to degrade TCE without aromatic induction may be useful for bioremediation of TCE-containing aquifers and groundwater. Thus, a simulated aquifer sediment system and groundwater microcosms were used to monitor the survival of G4 5223-PR1. The fate of G4 5223-PR1 in sediment was monitored by indirect immunofluorescence microscopy, a colony blot assay, and growth on selective medium. G4 5223-PR1 was detected immunologically by using a highly specific monoclonal antibody which reacted against the O-specific polysaccharide chain of the lipopolysaccharides of this organism. G4 5223-PR1 survived well in sterilized groundwater, although in nonsterile groundwater microcosms rapid decreases in the G4 5223-PR1 cell population were observed. Ten days after inoculation no G4 5223-PR1 cells could be detected by selective plating or immunofluorescence. G4 5223-PR1 survival was greater in a nonsterile aquifer sediment microcosm, although after 22 days of elution the number of G4 5223-PR1 cells was low. Our results demonstrate the utility of monoclonal antibody tracking methods and the importance of biotic interactions in determining the persistence of introduced microorganisms.