测定细菌竞争引起的抗菌活性的定性和定量方法。

Boris Taillefer, Marie M Grandjean, Julien Herrou, Donovan Robert, Tâm Mignot, Corinne Sebban-Kreuzer, Eric Cascales
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引用次数: 1

摘要

在环境中,细菌争夺生态位和资源;因此,它们进化出了各种各样的抗菌武器来摧毁竞争对手。目前用于抗菌活性评估的实验室技术通常是劳动密集型、低通量、昂贵和耗时的。典型的实验依赖于竞争后猎物细胞在选择性固体培养基上的菌落生长。在这里,我们提出了快速、廉价、互补的优化方案来定性和定量地测量抗菌活性。第一种方法是基于β-半乳糖苷酶的细胞不渗透显色底物的降解,β-半乳糖苷酶是一种在被攻击的报告菌株裂解过程中释放的细胞质酶。第二种方法依赖于被攻击细胞在竞争后达到规定光密度所需的滞后时间,这直接依赖于存活细胞的初始数量。第一种方法利用β-半乳糖苷酶的释放作为细菌裂解的代理。第二种方法是基于存活细胞的生长时间。两种方法的结合区分细胞死亡和裂解,细胞死亡不裂解,或存活到准裂解。方法对大肠埃希菌、铜绿假单胞菌、黄粘球菌等多种细菌进行优化。图形的概述。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Qualitative and Quantitative Methods to Measure Antibacterial Activity Resulting from Bacterial Competition.

In the environment, bacteria compete for niche occupancy and resources; they have, therefore, evolved a broad variety of antibacterial weapons to destroy competitors. Current laboratory techniques to evaluate antibacterial activity are usually labor intensive, low throughput, costly, and time consuming. Typical assays rely on the outgrowth of colonies of prey cells on selective solid media after competition. Here, we present fast, inexpensive, and complementary optimized protocols to qualitatively and quantitively measure antibacterial activity. The first method is based on the degradation of a cell-impermeable chromogenic substrate of the β-galactosidase, a cytoplasmic enzyme released during lysis of the attacked reporter strain. The second method relies on the lag time required for the attacked cells to reach a defined optical density after the competition, which is directly dependent on the initial number of surviving cells. Key features First method utilizes the release of β-galactosidase as a proxy for bacterial lysis. Second method is based on the growth timing of surviving cells. Combination of two methods discriminates between cell death and lysis, cell death without lysis, or survival to quasi-lysis. Methods optimized to various bacterial species such as Escherichia coli, Pseudomonas aeruginosa, and Myxococcus xanthus. Graphical overview.

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