Kubistova Lucie, Dvoracek Lukas, Tkadlec Jan, Melter Oto, Licha Irena
1 Department of Genetics and Microbiology, Faculty of Science, Charles University , Prague, Czech Republic .
2 Department of Medical Microbiology, 2nd Faculty of Medicine, Charles University , Prague, Czech Republic .
Microb Drug Resist. 2018 Jun;24(5):547-555. doi: 10.1089/mdr.2017.0064. Epub 2017 Aug 16.
The ability to form persisters has been observed in many microorganisms, including Staphylococcus aureus, mainly in the context of chronic infections and the pathogenicity of these microbes. In our research, we have demonstrated that salt or oxidative stress could play a role in the formation of S. aureus persisters outside the host's intracellular interface. We pre-exposed planktonic growing bacterial culture to an oxidative or salt stress and monitored the dynamics of persister formation after ciprofloxacin and gentamicin treatment. In parallel, using the quantitative PCR (qPCR) approach, we determined the expression level of the stress sigma factor SigB. The pre-exposure of bacteria to salt stress caused a 1-2.5 order of magnitude increase in persister formation in the bacterial population after antibiotic exposure, depending on the type and concentration of the antibiotic used. In contrast, oxidative stress only minimally influenced the formation of persisters, without correlation to the antibiotic type and concentration. We have demonstrated that both stress and antibiotic exposure increase the expression of sigB in bacterial populations from very early on. And that the expression level of sigB differs with the type of antibiotic and stress, but no correlation was observed between persister formation and sigB expression. The method used could be helpful in testing the ability that strains can have, to form persisters.
在包括金黄色葡萄球菌在内的许多微生物中都观察到了形成持留菌的能力,这主要发生在慢性感染以及这些微生物的致病性方面。在我们的研究中,我们已经证明盐胁迫或氧化应激可能在宿主细胞内界面之外的金黄色葡萄球菌持留菌形成中发挥作用。我们将浮游生长的细菌培养物预先暴露于氧化或盐胁迫下,并监测环丙沙星和庆大霉素处理后持留菌形成的动态变化。同时,我们使用定量PCR(qPCR)方法测定了应激σ因子SigB的表达水平。细菌预先暴露于盐胁迫下,抗生素暴露后细菌群体中持留菌的形成增加了1 - 2.5个数量级,这取决于所用抗生素的类型和浓度。相比之下,氧化应激仅对持留菌的形成产生极小的影响,且与抗生素类型和浓度无关。我们已经证明,应激和抗生素暴露从很早就增加了细菌群体中sigB的表达。并且sigB的表达水平因抗生素类型和应激而异,但在持留菌形成与sigB表达之间未观察到相关性。所使用的方法可能有助于测试菌株形成持留菌的能力。