次氯酸钠与酶联合对不锈钢表面细菌生物被膜的清除作用

Removal of bacterial biofilm from stainless steel surfaces by NaClO in combination with enzymes

定植于设备不同位置的细菌生物被膜是食品工业中应用清洁和消毒程序面临的最大挑战。该研究目的是通过人工培养生物被膜进行不同菌株生物被膜形成能力筛选,采用酶、次氯酸钠以及二者联合的方式对不锈钢试材上的成熟膜进行清除,通过平板计数、流式细胞仪等手段去探究不同处理方式的差异。结果表明,酶的抗生物被膜功效因生物被膜形成阶段的不同而体现出差异,在生物被膜形成期间,酶处理往往比对成熟生物被膜更有效,部分酶的参与会导致细菌产生更多的生物被膜基质。细菌计数显示,3种处理方式分别导致枯草芽孢杆菌(Bacillus subtilis)CICC 10900、金黄色葡萄球菌(Staphylococcus aureus)ATCC 6538细胞群减少0.64~3.08 lg CFU/cm^(2)、0.81~2.19 lg CFU/cm^(2);流式细胞仪结果表明,经处理后7.09%~12.04%、22.43%~23.45%、31.33%~37.78%的细胞受到损伤或死亡。使用酶制剂可以帮助次氯酸钠等二级制剂更有效地渗透生物被膜,增强对细菌细胞的清除功效。该研究结果可对食品工业在实施生物被膜清除程序时提供数据支撑和理论参考。

Bacterial biofilms colonizing different locations of equipment are the biggest challenge for the application of cleaning and disinfection procedures in the food industry.The purpose of this study was to screen the biofilm forming ability of different strains of bacteria by artificially culturing biofilms,using enzymes,NaClO,and a combination of both to remove mature films from stainless steel test materials,and to investigate the differences between the different treatments by means of plate counting and flow cytometry.Results showed that the anti-biofilm efficacy of enzymes varied depending on the stage of biofilm formation,with enzyme treatment tending to be more effective during biofilm formation than for mature biofilms,and partial enzyme involvement leading to the production of more biofilm substrates by bacteria.Bacterial counts showing that the three treatments resulted in reductions of 0.64 lg CFU/cm^(2) to 3.08 lg CFU/cm^(2) and 0.81 lg CFU/cm^(2) to 2.19 lg CFU/cm^(2) in Bacillus subtilis CICC 10900,Staphylococcus aureus ATCC 6538 cell populations,respectively.Flow cytometry results showed that 7.09%-12.04%,22.43%-23.45%,and 31.33%-37.78%of the cells were damaged or died after treatment.The use of enzymes can help secondary agents such as NaClO to penetrate the biofilm more effectively and enhance the efficacy of removal of bacterial cells.The results of this study can provide data support and theoretical reference for the food industry when implementing biofilm removal procedures.