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首页> 外文期刊>Applied Microbiology >Stress Resistance and Pathogenicity of Nonthermal-Plasma-Induced Viable-but-Nonculturable Staphylococcus aureus through Energy Suppression, Oxidative Stress Defense, and Immune-Escape Mechanisms
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Stress Resistance and Pathogenicity of Nonthermal-Plasma-Induced Viable-but-Nonculturable Staphylococcus aureus through Energy Suppression, Oxidative Stress Defense, and Immune-Escape Mechanisms

机译：通过能量抑制，氧化应激防御和免疫逃逸机制来抗血浆诱导的可行性但不培养的葡萄球菌的应力抗性和致胁迫性

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The occurrence of viable-but-nonculturable (VBNC) bacteria poses a potential risk to food safety due to failure in conventional colony detection. In this study, induction of VBNC Staphylococcus aureus was conducted by exposure to an atmospheric-pressure air dielectric barrier discharge-nonthermal-plasma (DBD-NTP) treatment with an applied energy of 8.1 kJ. The stress resistance profiles and pathogenicity of VBNC S. aureus were further evaluated. We found that VBNC S. aureus showed levels of tolerance of heat, acid, and osmosis challenges comparable to those shown by culturable S. aureus , while VBNC S. aureus exhibited enhanced resistance to oxidative and antibiotic stress, relating to the mechanisms of cellular energy depletion, antioxidant response initiation, and multidrug efflux pump upregulation. Regarding pathogenicity, NTP-induced VBNC S. aureus retained the capacity to infect the HeLa host cells. Compared with the culturable counterparts, VBNC S. aureus caused reduced immune responses (Toll-like receptor [TLR], nucleotide-binding oligomerization domain [NOD]) in HeLa cells, which was attributed to suppression of biosynthesis of the recognized surface ligands (e.g., peptidoglycan). Additionally, the proteomic analysis revealed that upregulation of several virulence factors (ClfB, SdrD, SCIN, SasH, etc.) could ensure that VBNC S. aureus would adhere to and internalize into host cells and avoid the host attack. The camouflaged mechanisms described above led to VBNC S. aureus causing less damage to the host cells, and their activity might result in longer intracellular persistence, posing potential risks during NTP processing.IMPORTANCE The consumer demand for freshness and nutrition has accelerated the development of mild decontamination technologies. The incomplete killing of nonthermal (NT) treatments might induce pathogens to enter into a viable-but-nonculturable (VBNC) status as a survival strategy. The use of nonthermal plasma (NTP) as a novel food decontamination technology received increased attention in food industry during recent decades. Our previous work confirmed that the foodborne pathogen S. aureus was induced into VBNC status in response to NTP exposure. This work further revealed the development of stress resistance and virulence retention of NTP-induced VBNC S. aureus through the mechanisms of energy suppression, oxidative stress defense, and immune escape. The data provide fundamental knowledge of the potential risks posed by NTP-induced VBNC S. aureus , which require further parameter optimization of the NTP process or combination with other techniques to avoid the occurrence of VBNC bacteria.
机译：活性但不可培养的（VBNC）细菌的发生潜在的冒险风险导致常规殖民地检测失败引起的食品安全性。在该研究中，通过暴露于8.1kj的施加能量的施加能量，通过暴露于大气压空气介电阻挡放电 - 非热血浆（DBD-NTP）处理来进行VBNC金黄色葡萄球菌的诱导。进一步评估VBNC S.UUREUS的应力抗性分布和致病性。我们发现VBNC S.UUREUS显示出与培养的S.金黄色葡萄球菌所示的热量，酸和渗透挑战的耐受性差异，而VBNC S.UUREUS表现出对氧化和抗生素应激的抗性增强，与细胞能量的机制有关耗尽，抗氧化反应起始和多药渗透泵上调。关于致病性，NTP诱导的VBNC S.UUREUS保留了感染HELA宿主细胞的能力。与培养对应物相比，VBNC S.UUREUS在HELA细胞中引起了降低的免疫应答（诸如受体[TLR]，核苷酸结合寡聚化结构域[NOD]），其归因于抑制识别的表面配体的生物合成（例如，肽聚糖）。另外，蛋白质组学分析表明，若干毒力因子（CLFB，SDRD，Scin，SASH等）的上调可以确保VBNC S.UUREUS将粘附并内化到宿主细胞中并避免宿主攻击。上述伪装机制导致VBNC S.UUREUS导致宿主细胞的损害较少，并且它们的活动可能导致较长的细胞内持久性，在NTP处理期间构成潜在风险。分为消费者对新鲜度和营养的需求加速了轻度的发展净化技术。不完全杀害非热（NT）治疗可能会诱导病原体，以进入可行的但不可渗透的（VBNC）状态作为生存策略。在近几十年来，使用非热等离子体（NTP）作为新型食品去污技术得到了食品行业的关注。我们以前的作品证实，响应于NTP暴露，诱导Footborne病原体S.金黄色葡萄球菌诱导VBNC状态。这项工作进一步揭示了通过能量抑制，氧化胁迫防御和免疫逃逸的机制发展NTP诱导的VBNC S.金黄色葡萄球菌的胁迫抗性和毒力保留。数据提供了NTP诱导的VBNC S.UUREUS所带来的潜在风险的基本知识，这需要进一步参数优化NTP过程或与其他技术组合，以避免VBNC细菌的发生。

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