Berney Michael, Weilenmann Hans-Ulrich, Ihssen Julian, Bassin Claudio, Egli Thomas
Department of Environmental Microbiology, Swiss Federal Institute of Aquatic Science and Technology, Uberlandstrasse 133, P.O. Box 611, CH-8600 Dübendorf, Switzerland.
Appl Environ Microbiol. 2006 Apr;72(4):2586-93. doi: 10.1128/AEM.72.4.2586-2593.2006.
Knowledge about the sensitivity of the test organism is essential for the evaluation of any disinfection method. In this work we show that sensitivity of Escherichia coli MG1655 to three physical stresses (mild heat, UVA light, and sunlight) that are relevant in the disinfection of drinking water with solar radiation is determined by the specific growth rate of the culture. Batch- and chemostat-cultivated cells from cultures with similar specific growth rates showed similar stress sensitivities. Generally, fast-growing cells were more sensitive to the stresses than slow-growing cells. For example, slow-growing chemostat-cultivated cells (D = 0.08 h(-1)) and stationary-phase bacteria from batch culture that were exposed to mild heat had very similar T(90) (time until 90% of the population is inactivated) values (T(90, chemostat) = 2.66 h; T(90, batch) = 2.62 h), whereas T(90) for cells growing at a mu of 0.9 h(-1) was 0.2 h. We present evidence that the stress sensitivity of E. coli is correlated with the intracellular level of the alternative sigma factor RpoS. This is also supported by the fact that E. coli rpoS mutant cells were more stress sensitive than the parent strain by factors of 4.9 (mild heat), 5.3 (UVA light), and 4.1 (sunlight). Furthermore, modeling of inactivation curves with GInaFiT revealed that the shape of inactivation curves changed depending on the specific growth rate. Inactivation curves of cells from fast-growing cultures (mu = 1.0 h(-1)) that were irradiated with UVA light showed a tailing effect, while for slow-growing cultures (mu = 0.3 h(-1)), inactivation curves with shoulders were obtained. Our findings emphasize the need for accurate reporting of specific growth rates and detailed culture conditions in disinfection studies to allow comparison of data from different studies and laboratories and sound interpretation of the data obtained.
了解测试微生物的敏感性对于评估任何消毒方法至关重要。在本研究中,我们表明,大肠杆菌MG1655对饮用水太阳能辐射消毒中相关的三种物理应激(温和加热、紫外线A光和阳光)的敏感性由培养物的比生长速率决定。来自具有相似比生长速率的培养物的分批培养和恒化器培养细胞表现出相似的应激敏感性。一般来说,快速生长的细胞比缓慢生长的细胞对应激更敏感。例如,缓慢生长的恒化器培养细胞(D = 0.08 h(-1))和分批培养中处于稳定期的细菌在受到温和加热时具有非常相似的T(90)(直至90%的群体失活的时间)值(T(90, 恒化器) = 2.66 h;T(90, 分批) = 2.62 h),而比生长速率为0.9 h(-1)的细胞的T(90)为0.2 h。我们提供的证据表明,大肠杆菌的应激敏感性与替代sigma因子RpoS的细胞内水平相关。这也得到了以下事实的支持:大肠杆菌rpoS突变细胞对应激的敏感性比亲本菌株高4.9倍(温和加热)、5.3倍(紫外线A光)和4.1倍(阳光)。此外,用GInaFiT对失活曲线进行建模表明,失活曲线的形状根据比生长速率而变化。用紫外线A光照射的快速生长培养物(μ = 1.0 h(-1))的细胞的失活曲线显示出拖尾效应,而对于缓慢生长的培养物(μ = 0.3 h(-1)),则获得了带有肩部的失活曲线。我们的研究结果强调,在消毒研究中需要准确报告比生长速率和详细的培养条件,以便能够比较来自不同研究和实验室的数据,并对所获得的数据进行合理的解释。