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Effects of Contact Time, Pressure, Percent Relative Humidity (%RH), and Material Type on Listeria Biofilm Adhesive Strength at a Cellular Level Using Atomic Force Microscopy (AFM)

机译:接触时间,压力,相对湿度百分比(%RH)和材料类型对使用原子力显微镜(AFM)在细胞水平上对李斯特菌生物膜粘附强度的影响

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Whereas the transfer of Listeria from surfaces to foods and vice versa has been well documented, little is known about the mechanism of bacterial transfer. The objective of this work is to gain a better understanding of the forces involved in listerial biofilms adhesion using atomic force microscopy (AFM). L. monocytogenes Scott A was grown as biofilms on stainless steel surfaces by inoculating stainless steel coupons with Listeria and incubating the coupons for 48 h at 32 ℃ with a diluted 1:20 tryptic soy broth. After growth, biofilms were equilibrated over saturated salt solutions at a constant relative humidity (%RH) before measurement of adhesion forces using AFM. The effects of contact time, loading force, and biofilm relative humidity (%RH) suggested that neither contact time, loading force nor biofilm %RH had a significant effect on biofilm adhesiveness at a cellular level (P>0.05). In a second set of experiments, the influence of material type on biofilm adhesiveness was evaluated using two different colloidal probes (SiO_2 and polyethylene). Results showed that the maximum pull-off force andrnretraction work needed to retract the cantilever for glass (-85.42 nN and 1.610~(-15) J, respectively) were significantly lower than those of polyethylene (-113.38 nN and 2.7× 10~(-15) J, respectively; P<0.001). The results of this study suggest that Listeria biofilms adhere more strongly to hydrophobic surfaces than hydrophilic surfaces when measured at a cellular level. These results provide important insights that could lead to new ways to remediate and avoid listerial biofilm formation in the food industry.
机译:尽管已经充分记录了李斯特菌从表面到食物的转移,反之亦然,但对细菌转移的机制知之甚少。这项工作的目的是要使用原子力显微镜(AFM)更好地了解涉及李斯特菌生物膜粘附的力。单核细胞增生李斯特氏菌斯科特A通过在李斯特氏菌中接种不锈钢试片,并在32℃下用稀释的1:20胰蛋白酶大豆肉汤孵育48小时,从而在不锈钢表面上生长成生物膜。生长后,在使用AFM测量粘附力之前,将生物膜在恒定的相对湿度(%RH)上的饱和盐溶液中平衡。接触时间,负载力和生物膜相对湿度(%RH)的影响表明,接触时间,负载力和生物膜%RH在细胞水平上均对生物膜粘附性没有显着影响(P> 0.05)。在第二组实验中,使用两种不同的胶体探针(SiO_2和聚乙烯)评估了材料类型对生物膜粘附性的影响。结果表明,使玻璃悬臂缩回所需的最大拉力和最大拉拔功(分别为-85.42 nN和1.610〜(-15)J)显着低于聚乙烯(-113.38 nN和2.7×10〜( -15)J; P <0.001)。这项研究的结果表明,在细胞水平上测量,利斯特氏菌生物膜比疏水性表面更牢固地粘附于疏水性表面。这些结果提供了重要的见解,可能会导致新的方法来补救和避免食品工业中李斯特菌生物膜的形成。

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