CSIRO Food and Nutritional Sciences, Coopers Plains, Queensland 4108, Australia.
Appl Environ Microbiol. 2011 Oct;77(20):7339-44. doi: 10.1128/AEM.00277-11. Epub 2011 Aug 19.
The role of curli expression in attachment of Escherichia coli O157:H7 to glass, Teflon, and stainless steel (SS) was investigated through the creation of csgA knockout mutants in two isolates of E. coli O157:H7. Attachment assays using epifluorescence microscopy and measurements of the force of adhesion of bacterial cells to the substrates using atomic force microscopy (AFM) force mapping were used to determine differences in attachment between wild-type (wt) and csgA-negative (ΔcsgA) strains following growth in four different media. The hydrophobicity of the cells was determined using contact angle measurements (CAM) and bacterial adhesion to hydrocarbons (BATH). The attachment assay results indicated that ΔcsgA strains attached to glass, Teflon, and SS surfaces in significantly different numbers than their wt counterparts in a growth medium-dependent fashion (P < 0.05). However, no clear correlation was seen between attachment numbers, surface type, or growth medium. No correlation was seen between BATH and CAM results (R(2) < 0.70). Hydrophobicity differed between the wt and ΔcsgA in some cases in a growth medium- and method-dependent fashion (P < 0.05). AFM force mapping revealed no significant difference in the forces of adhesion to glass and SS surfaces between wt and ΔcsgA strains (P > 0.05) but a significantly greater force of adhesion to Teflon for one of the two wt strains than for its ΔcsgA counterpart (P < 0.05). This study shows that CsgA production by E. coli O157:H7 may alter attachment behavior in some environments; however, further investigation is required in order to determine the exact relationship between CsgA production and attachment to abiotic surfaces.
通过在两种大肠杆菌 O157:H7 分离株中创建 csgA 敲除突变体,研究了卷曲表达在大肠杆菌 O157:H7 附着到玻璃、特氟龙和不锈钢 (SS) 上的作用。使用荧光显微镜的附着实验和原子力显微镜 (AFM) 力映射测量细菌细胞对基底的附着力,以确定在四种不同培养基中生长后,野生型 (wt) 和 csgA 阴性 (ΔcsgA) 菌株之间的附着差异。使用接触角测量 (CAM) 和细菌对烃类的粘附 (BATH) 来确定细胞的疏水性。附着实验结果表明,ΔcsgA 菌株以依赖于生长培养基的方式比其 wt 对应物附着到玻璃、特氟龙和 SS 表面的数量明显不同 (P < 0.05)。然而,在附着数量、表面类型和生长培养基之间没有看到明显的相关性。在 BATH 和 CAM 结果之间没有看到相关性 (R(2) < 0.70)。在某些情况下,在依赖于生长培养基和方法的情况下,wt 和 ΔcsgA 之间的疏水性存在差异 (P < 0.05)。AFM 力映射显示 wt 和 ΔcsgA 菌株对玻璃和 SS 表面的附着力没有显著差异 (P > 0.05),但其中一种 wt 菌株对特氟龙的附着力明显大于其 ΔcsgA 对应物 (P < 0.05)。这项研究表明,大肠杆菌 O157:H7 中 CsgA 的产生可能会改变某些环境中的附着行为;然而,需要进一步的研究来确定 CsgA 产生与非生物表面附着之间的确切关系。