Bacterial Cell Surface Damage Due to Centrifugal Compaction
Bacterial Cell Surface Damage Due to Centrifugal Compaction
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DOI：
10.1128/aem.06780-11
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发表时间：
2012-01-01
影响因子：
4.4
通讯作者：
Busscher, Henk J.
中科院分区：
文献类型：
--
作者：
Peterson, Brandon W.;Sharma, Prashant K.;Busscher, Henk J.
已知离心性损伤会改变细菌细胞的表面性质和内部结构，包括DNA。关于离心法造成的细菌损伤的研究很少，因为很难将离心机的速度和容器的几何形状与所造成的损伤联系起来。在这里，我们提供了一种简单、通用的分析方法，用于描述离心过程中细菌的紧实度，基于提出的离心系数C。C的值可以与不同的细菌细胞表面属性相关。改变离心容器的几何形状或改变离心速度会显著改变C的值。金黄色葡萄球菌ATCC 12600在玻璃表面的初始沉积速率随C的增加而指数衰减，从4,217 cm(-2)到1,478 cm(-2)S(-1)，而Zeta电位在-15 mV左右的葡萄球菌所占比例从97%下降到58%。这些表面敏感参数被独立使用来推导出临界离心系数(0.040)，超过该系数后，由于细胞表面损伤，离心力被认为会影响表面敏感实验的结果。在84%的病例中，临界离心力系数可以成功地预测葡萄球菌细胞表面损伤，即初始沉积率或Zeta电位分布的显著变化，而离心速度只能预测58%的病例的损伤。此外，在一系列实验中，将离心系数控制在狭窄的范围内，与重复实验的固定速度离心法相比，初始葡萄球菌沉积率的标准偏差小43%。
Centrifugal damage has been known to alter bacterial cell surface properties and interior structures, including DNA. Very few studies exist on bacterial damage caused by centrifugation because of the difficulty in relating centrifugation speed and container geometry to the damage caused. Here, we provide a simple, versatile method of analysis for describing the compaction of bacteria during centrifugation based on a proposed centrifugation coefficient, C. Values of C can be related to different bacterial cell surface properties. Changing the geometry of the centrifugation container or centrifugation speeds changed the value of C significantly. Initial deposition rates of Staphylococcus aureus ATCC 12600 to a glass surface decayed exponentially from 4,217 to 1,478 cm(-2) s(-1) with increasing C, while the proportion of staphylococci with a zeta potential of around -15 mV decreased from 97 to 58%. These surface-sensitive parameters were used independently to derive a critical centrifugation coefficient (0.040), above which centrifugation was considered to impact the outcome of surface-sensitive experiments due to cell surface damage. The critical centrifugation coefficient could successfully predict staphylococcal cell surface damage, i.e., a significant change in initial deposition rate or zeta potential distribution, in 84% of all cases included here, whereas the centrifugation speed could predict damage in only 58% of all cases. Moreover, controlling the centrifugation coefficient within narrow limits over a series of experiments yielded 43% smaller standard deviations in initial staphylococcal deposition rates than with centrifugation at fixed speeds for replicate experiments.