Department of Food Science and Agricultural Chemistry, Faculty of Agricultural and Environmental Sciences, McGill University, Sainte-Anne-de-Bellevue, Quebec, Canada.
Appl Environ Microbiol. 2023 May 31;89(5):e0009623. doi: 10.1128/aem.00096-23. Epub 2023 Apr 17.
Many bacteria can become viable but nonculturable (VBNC) in response to stressors commonly identified in agrifood systems. Campylobacter is able to enter the VBNC state to evade unfavorable environmental conditions, but how food processing can induce Campylobacter jejuni to enter this state and the potential role of foods in inducing the VBNC state in C. jejuni remains largely unknown. In this study, the culturability and viability of C. jejuni cells were investigated under chlorine treatment (25 ppm), aerobic stress (atmospheric condition), and low-temperature (4°C) conditions that mimicked food processing. In addition, the behaviors of C. jejuni cells in ultrahigh-temperature (UHT) and pasteurized milk were also monitored during refrigerated storage. The numbers of viable and culturable C. jejuni cells in both the pure bacterial culture and food matrices were separately determined by propidium monoazide (PMA)-quantitative PCR (qPCR) and plating assay. The C. jejuni cells lost their culturability but partially retained their viability (1% to 10%) once mixed with chlorine. In comparison, ~10% of C. jejuni cells were induced to enter the VBNC state after 24 h and 20 days under aerobic and low-temperature conditions, respectively. The viability of the C. jejuni cells remained stable during the induction process in UHT (>10%) and pasteurized (>10%) milk. The number of culturable C. jejuni cells decreased quickly in pasteurized milk, but culturable cells could still be detected in the end (day 21). In contrast, the number of culturable C. jejuni cells slowly decreased, and they became undetectable after >42 days in UHT milk. The C. jejuni cells responded differently to various stress conditions and survived in high numbers in the VBNC state in agrifood systems. The VBNC state of pathogens can pose risks to food safety and public health because the pathogens cannot be detected using conventional microbiological culture-based methods but can resuscitate under favorable conditions to develop virulence. As a leading cause of human gastroenteritis worldwide, C. jejuni can enter the VBNC state to survive in the environment and food-processing chain with high prevalence. In this study, the effect of food-processing conditions and food products on the development of VBNC state in C. jejuni was investigated, providing a better understanding of the interaction between C. jejuni and the agroecosystem. The knowledge elicited from this study can aid in developing novel intervention strategies to reduce the food safety risks associated with this microbe.
许多细菌能够在应对农业食品系统中常见的应激源时变为存活但不可培养状态(VBNC)。空肠弯曲菌能够进入 VBNC 状态以逃避不利的环境条件,但食品加工如何诱导空肠弯曲菌进入这种状态以及食物在诱导空肠弯曲菌进入 VBNC 状态中的潜在作用在很大程度上仍不清楚。在这项研究中,研究了氯处理(25 ppm)、需氧应激(大气条件)和模拟食品加工的低温(4°C)条件下空肠弯曲菌细胞的可培养性和生存能力。此外,还监测了超高温(UHT)和巴氏杀菌乳中空肠弯曲菌细胞在冷藏储存过程中的行为。通过叠氮溴化丙啶(PMA)定量 PCR(qPCR)和平板计数法分别确定了纯细菌培养物和食品基质中存活和可培养的空肠弯曲菌细胞的数量。当与氯混合时,空肠弯曲菌细胞失去了可培养性,但部分保留了其生存能力(1%至 10%)。相比之下,在需氧和低温条件下分别经过 24 小时和 20 天处理后,约 10%的空肠弯曲菌细胞被诱导进入 VBNC 状态。在 UHT(>10%)和巴氏杀菌(>10%)乳中,空肠弯曲菌细胞的生存能力在诱导过程中保持稳定。巴氏杀菌乳中可培养的空肠弯曲菌细胞数量迅速减少,但在实验结束时仍能检测到可培养细胞(第 21 天)。相比之下,UHT 乳中可培养的空肠弯曲菌细胞数量缓慢减少,>42 天后无法检测到。空肠弯曲菌细胞对各种应激条件的反应不同,在农业食品系统中以大量数量存在于 VBNC 状态下。由于病原体不能通过常规的微生物培养基于方法检测到,但可以在有利条件下复苏以产生毒力,因此病原体的 VBNC 状态会对食品安全和公共健康构成威胁。空肠弯曲菌是全球人类肠胃炎的主要原因之一,它可以在环境和食品加工链中以高流行率进入 VBNC 状态生存。在这项研究中,研究了食品加工条件和食品产品对空肠弯曲菌 VBNC 状态发展的影响,为更好地了解空肠弯曲菌与农业生态系统之间的相互作用提供了依据。从这项研究中获得的知识可以帮助制定新的干预策略,以降低与这种微生物相关的食品安全风险。