Department of Food and Nutrition, Chung-Ang University, Anseong-si, Republic of Korea.
Lett Appl Microbiol. 2021 Jun;72(6):741-749. doi: 10.1111/lam.13468. Epub 2021 Mar 6.
In the present study, Escherichia coli O157:H7 and Salmonella enterica serovar Enteritidis were transferred into Luria-Bertani medium without NaCl (LBWS) and adjusted to various pHs (4, 5, 6 and 7) with lactic acid containing 0·75, 5, 10 and 30% NaCl, and stored at 25°C until the bacterial populations reached below detectable levels on tryptic soy agar (TSA). Although E. coli O157:H7 and S. Enteritidis did not grow on TSA when incubated in LBWS with 30% NaCl for 35 and 7 days, more than 60 and 70% of the bacterial cells were shown to be viable via fluorescent staining with SYTO9 and propidium iodide (PI), respectively, suggesting that a number of cells could be induced into the viable but nonculturable (VBNC) state. These bacteria that were induced into a VBNC state were transferred to a newly prepared tryptic soy broth (TSB) and then incubated at 37°C for several days. After more than 7 days, E. coli O157:H7 and S. Enteritidis regained their culturability. We, therefore, suggest that E. coli O157:H7 and S. Enteritidis entered the VBNC state under the adverse condition of higher salt concentrations and were revived when these conditions were reversed.
在本研究中,将大肠杆菌 O157:H7 和肠炎沙门氏菌血清型肠炎传入不含 NaCl 的 Luria-Bertani 培养基(LBWS)中,并使用含有 0.75、5、10 和 30% NaCl 的乳酸将 pH 值调节至 4、5、6 和 7,并在 25°C 下储存,直到细菌数量在胰蛋白胨大豆琼脂(TSA)上降至无法检测的水平。尽管在含有 30% NaCl 的 LBWS 中孵育 35 和 7 天时,大肠杆菌 O157:H7 和肠炎沙门氏菌在 TSA 上无法生长,但通过使用 SYTO9 和碘化丙啶(PI)进行荧光染色,分别显示超过 60%和 70%的细菌细胞仍具有活力,表明有许多细胞可能被诱导进入活但非可培养(VBNC)状态。这些被诱导进入 VBNC 状态的细菌被转移到新制备的胰蛋白胨大豆肉汤(TSB)中,然后在 37°C 下孵育数天。7 天后,大肠杆菌 O157:H7 和肠炎沙门氏菌恢复了可培养性。因此,我们认为大肠杆菌 O157:H7 和肠炎沙门氏菌在高盐浓度的不利条件下进入 VBNC 状态,当这些条件逆转时,它们会复苏。