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酸适应增强了血清型鼠伤寒沙门氏菌和 O157:H7 对 222 纳米氪-氯 excilamp 处理的抗性。

Increased Resistance of Serovar Typhimurium and O157:H7 to 222-Nanometer Krypton-Chlorine Excilamp Treatment by Acid Adaptation.

机构信息

Department of Agricultural Biotechnology, Center for Food and Bioconvergence, Research Institute for Agricultural and Life Sciences, Seoul National University, Seoul, Republic of Korea.

Institutes of Green-bio Science & Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Republic of Korea.

出版信息

Appl Environ Microbiol. 2019 Mar 6;85(6). doi: 10.1128/AEM.02221-18. Print 2019 Mar 15.

Abstract

In this study, we examined the change in resistance of serovar Typhimurium and O157:H7 to 222-nm krypton-chlorine (KrCl) excilamp treatment as influenced by acid adaptation and identified a mechanism of resistance change. In addition, we measured changes in apple juice quality indicators, such as color, total phenols, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, during treatment. Non-acid-adapted and acid-adapted pathogens were induced by growing the cells in tryptic soy broth without dextrose (TSB w/o D) at pH 7.3 and in TSB w/o D at pH 5.0 (adjusted with HCl), respectively. For the KrCl excilamp treatment, acid-adapted pathogens exhibited significantly (0.05) higher D values, which indicate dosages required to achieve a 5-log reduction, than those for non-acid-adapted pathogens in both commercially clarified apple juice and phosphate-buffered saline (PBS), and the pathogens in the juice showed significantly (0.05) higher D values than those for pathogens in PBS because of the UV-absorbing characteristics of apple juice. Through mechanism identification, it was found that the generation of lipid peroxidation in the cell membrane, inducing cell membrane destruction, was significantly (0.05) lower in acid-adapted cells than in non-acid-adapted cells for the same amount of reactive oxygen species (ROS) generated at the same dose because the ratio of unsaturated to saturated fatty acids (USFA/SFA) in the cell membrane was significantly (0.05) decreased as a result of acid adaptation. Treated apple juice showed no significant (0.05) difference in quality indicators compared to those of untreated controls during treatment at 1,773 mJ/cm There is a need for novel, mercury-free UV lamp technology to replace germicidal lamps containing harmful mercury, which are routinely utilized for UV pasteurization of apple juice. In addition, consideration of the changes in response to antimicrobial treatments that may occur when pathogens are adapted to the acid in an apple juice matrix is critical to the practical application of this technology. Based on this, an investigation using 222-nm KrCl excilamp technology, an attractive alternative to mercury lamps, was conducted. Our study demonstrated increased resistance to 222-nm KrCl excilamp treatment as pathogens adapted to acids, and this was due to changes in reactivity to ROS with changes in the fatty acid composition of the cell membrane. Despite increased resistance, the 222-nm KrCl excilamp achieved pathogen reductions of 5 log or more at laboratory scale without affecting apple juice quality. These results provide valuable baseline data for application of 222-nm KrCl excilamps in the apple juice industry.

摘要

在这项研究中,我们研究了酸适应对肠炎沙门氏菌和 O157:H7 对 222nm 氪-氯(KrCl)准分子灯处理的耐药性变化,并确定了耐药性变化的机制。此外,我们测量了处理过程中苹果汁质量指标的变化,如颜色、总酚和 2,2-二苯基-1-苦基肼(DPPH)自由基清除活性。非酸适应和酸适应的病原体分别通过在 pH7.3 的无葡萄糖胰蛋白酶大豆肉汤(TSB w/o D)和 pH5.0 的 TSB w/o D(用 HCl 调节)中生长细胞来诱导。对于 KrCl 准分子灯处理,酸适应的病原体在商业澄清苹果汁和磷酸盐缓冲盐水(PBS)中表现出显著(0.05)更高的 D 值,这表明达到 5 对数减少所需的剂量,而非酸适应的病原体;由于苹果汁的紫外线吸收特性,果汁中的病原体表现出显著(0.05)更高的 D 值。通过机制鉴定发现,在相同剂量下产生相同数量的活性氧(ROS)时,酸适应细胞中细胞膜的脂质过氧化生成,导致细胞膜破坏的程度显著(0.05)低于非酸适应细胞,因为细胞膜中不饱和脂肪酸与饱和脂肪酸的比例(USFA/SFA)显著(0.05)降低。与未经处理的对照相比,处理过的苹果汁在 1773mJ/cm2 下处理时,其质量指标没有显著差异(0.05)。需要开发新型的无汞紫外线灯技术来替代含有有害汞的杀菌灯,这些灯通常用于苹果汁的紫外线巴氏杀菌。此外,考虑到当病原体适应苹果汁基质中的酸时,对抗菌处理的反应可能发生的变化,对于这项技术的实际应用至关重要。基于此,我们使用 222nm KrCl 准分子灯技术进行了研究,该技术是汞灯的一种有吸引力的替代品。我们的研究表明,病原体适应酸后对 222nm KrCl 准分子灯处理的耐药性增加,这是由于细胞膜脂肪酸组成的变化导致对 ROS 的反应性发生变化。尽管耐药性增加,但 222nm KrCl 准分子灯在实验室规模上仍能达到 5 个对数级或更多的病原体减少,而不会影响苹果汁的质量。这些结果为在苹果汁行业应用 222nm KrCl 准分子灯提供了有价值的基线数据。

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