Formation and resuscitation of viable but non-culturable Campylobacter in agri-food industry

食品加工环境和食品中存在和复苏的活的但不可培养的（VBNC）病原体对公众健康构成重大风险。VBNC病原体可以通过常规培养方法逃避检测，但保持毒力，并且在复苏时有可能引起食源性疾病。空肠弯曲菌是引起人类胃肠道疾病的主要细菌，但其形成和复苏仍需进一步研究。空肠及其在食品工业中的控制措施尚未得到很好的研究。本研究的目的是探讨C.空肠在食品加工和储存相关环境以及食品中的应用。在C.空肠VBNC的形成，我们将监测C.空肠人口的过渡期间，从正常的可培养状态的VBNC状态下的各种压力条件下，模仿食品加工和保存。然后，我们将应用高级基础设施（例如，全细胞指纹谱和RNA测序分析）来表征C.空肠细胞进入VBNC状态。此外，我们还将开发一种与激光镊子拉曼光谱联用的微流控“芯片实验室”装置，用于对纯VBNC单分子的同时鉴定和分离。来自不同细菌亚群的混合物的空肠细胞（即，健康细胞、受损细胞和死细胞），然后使用单细胞转录组测序进行表征。在C.空肠VBNC复苏，我们将用代谢组学方法分析能促进VBNC复苏的农产品。空肠细胞，并确定潜在的复苏促进分子的食物。然后我们将研究复苏的C.利用RNA测序和蛋白质组学研究空肠细胞。最后，我们将制定降低正常和VBNC C的干预策略。环境和食物系统中的空肠。本研究结果可为VBNC C的快速、灵敏、易操作的检测和表征方法的发展提供参考。空肠弯曲菌在食品工业中的应用，以及减少弯曲菌在环境和农产品中流行的缓解战略。

The presence and resuscitation of viable but non-culturable (VBNC) pathogens in the food processing environment and food products pose a significant risk to the public health. VBNC pathogens can evade detection by the conventional culturing methods, but maintain virulence and have the potential to cause foodborne illnesses when resuscitated. Campylobacter jejuni is a leading bacterial cause of human gastrointestinal disease worldwide, but the formation and resuscitation of VBNC C. jejuni and its control measures in the food industry have not been well studied. The aim of this proposal is to investigate the formation and resuscitation of C. jejuni in food processing and storage-related environment as well as in food products. In the context of C. jejuni VBNC formation, we will monitor the dynamics of C. jejuni population during the transition from the normal culturable state to the VBNC state under various stress conditions that mimic food processing and preservation. We will then apply advanced infrastructures (e.g., whole cell fingerprinting spectroscopy and RNA-sequencing analysis) to characterize the biochemical and transcriptional profiles of C. jejuni cells induced into the VBNC state. Moreover, we will develop microfluidic “lab-on-a-chip” device coupled with laser tweezers Raman spectroscopy for the simultaneous identification and separation of pure single VBNC C. jejuni cells from a mixture of different bacterial subpopulations (i.e., healthy cells, injured cells, and dead cells), followed by characterization using single-cell transcriptome sequencing. In the context of C. jejuni VBNC resuscitation, we will use metabolomics to analyze agri-food products that can promote the resuscitation of VBNC C. jejuni cells and identify the potential resuscitation-promoting molecules in foods. We will then study the transcriptional and translational profiles of resuscitated C. jejuni cells using RNA-sequencing and proteomics. Finally, we will develop intervention strategies for the reduction of normal and VBNC C. jejuni in the environment and food system. The research outcome can provide an insight to the development of rapid, sensitive and easy-to-operate methods for the detection and characterization of VBNC C. jejuni in the food industry, and mitigation strategies to reduce the prevalence of Campylobacter in the environment and agri-foods.