Department of Food Hygiene and Technology, Veterinary Faculty, University of León, 24071 León, Spain.
Appl Environ Microbiol. 2010 Nov;76(22):7598-607. doi: 10.1128/AEM.00312-10. Epub 2010 Sep 17.
The effect of exposure to acid (pH 2.5), alkaline (pH 11.0), heat (55°C), and oxidative (40 mM H₂O₂) lethal conditions on the ultrastructure and global chemical composition of Salmonella enterica serovar Typhimurium CECT 443 cells was studied using transmission electron microscopy and Fourier transform infrared spectroscopy (FT-IR) combined with multivariate statistical methods (hierarchical cluster analysis and factor analysis). Infrared spectra exhibited marked differences in the five spectral regions for all conditions tested compared to those of nontreated control cells, which suggests the existence of a complex bacterial stress response in which modifications in a wide variety of cellular compounds are involved. The visible spectral changes observed in all of the spectral regions, together with ultrastructural changes observed by transmission electron microscopy and data obtained from membrane integrity tests, indicate the existence of membrane damage or alterations in membrane composition after heat, acid, alkaline, and oxidative treatments. Results obtained in this study indicate the potential of FT-IR spectroscopy to discriminate between intact and injured bacterial cells and between treatment technologies, and they show the adequacy of this technique to study the molecular aspects of bacterial stress response.
采用透射电子显微镜和傅里叶变换红外光谱(FT-IR)结合多元统计方法(层次聚类分析和因子分析)研究了暴露于酸(pH 2.5)、碱(pH 11.0)、热(55°C)和氧化(40 mM H₂O₂)致死条件对鼠伤寒沙门氏菌 CECT 443 细胞超微结构和整体化学成分的影响。与未经处理的对照细胞相比,所有测试条件下的红外光谱在五个光谱区域均显示出明显差异,这表明存在复杂的细菌应激反应,涉及多种细胞化合物的修饰。在所有光谱区域都观察到可见光谱变化,以及透射电子显微镜观察到的超微结构变化和膜完整性测试获得的数据表明,在热、酸、碱和氧化处理后存在膜损伤或膜成分改变。本研究结果表明 FT-IR 光谱有潜力区分完整和受损的细菌细胞以及不同的处理技术,并且表明该技术足以研究细菌应激反应的分子方面。