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在不同温度、低营养和高 NaCl 浓度条件下,处于活的非可培养状态下的李斯特菌的形成和恢复。

Formation and recovery of Listeria monocytogenes in viable but nonculturable state under different temperatures combined with low nutrition and high NaCl concentration.

机构信息

School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.

Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs, 299 Mian Bei Rd., Shanghai 201210, China.

出版信息

Food Res Int. 2024 Sep;192:114774. doi: 10.1016/j.foodres.2024.114774. Epub 2024 Jul 25.

Abstract

The viable but nonculturable (VBNC) state occurs when bacteria lose their ability to grow and multiply on conventional media when stressed by adverse environmental factors, but they remain active and can revive under certain conditions, posing a food safety risk. In this study, the VBNC state of Listeria monocytogenes was induced with different temperatures combined with low nutrient conditions; the VBNC state of L. monocytogenes was confirmed in conjunction with the housekeeping gene abcZ using a molecular biology assay (PMA-qPCR) to calculate the viable bacterial count; The resuscitation conditions for the VBNC state of L. monocytogenes were investigated utilizing various nutrients in the culture medium and pasteurized milk. Four strains of L. monocytogenes reached the VBNC stage after 14, 21, 21, and 35 days at 20°C with 20% (or 30%) NaCl. Resuscitation studies indicate that Trypticase Soy Broth (TSB) combined with Tween 80 and sodium pyruvate is more effective for resuscitation. The Chinese national standard technology GB 4789.30-2016 was used to inoculate lettuce, chicken, and pasteurized milk with L. monocytogenes ATCC 19115 VBNC state. This research has significant implications for commercial food processing, long-term storage, disinfection, disease prevention, and control.

摘要

当细菌在受到不利环境因素的压力时,失去在常规培养基上生长和繁殖的能力,但仍保持活跃并在某些条件下能够复苏时,就会出现活的但非可培养状态(VBNC)。在这项研究中,通过不同温度结合低营养条件诱导单核细胞增生李斯特菌进入 VBNC 状态;使用分子生物学检测(PMA-qPCR)结合管家基因 abcZ 来确认单核细胞增生李斯特菌的 VBNC 状态,以计算活菌数;通过在培养基和巴氏杀菌乳中添加各种营养物质来研究单核细胞增生李斯特菌 VBNC 状态的复苏条件。在 20°C 下,用 20%(或 30%)NaCl 处理 4 株单核细胞增生李斯特菌 14、21、21 和 35 天,达到 VBNC 阶段。复苏研究表明,Trypticase Soy Broth(TSB)与 Tween 80 和丙酮酸钠联合使用更有利于复苏。本研究采用中国国家标准技术 GB 4789.30-2016,用单核细胞增生李斯特菌 ATCC 19115 VBNC 状态接种生菜、鸡肉和巴氏杀菌乳。这项研究对于商业食品加工、长期储存、消毒、疾病预防和控制具有重要意义。

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