金黄色葡萄球菌活的非可培养状态的诱导和复苏

刘德欣; 刘佩莹; 陈颖翘; 田娟; 王丽; 钟青萍

doi:10.13386/j.issn1002-0306.2014.03.012

金黄色葡萄球菌活的非可培养状态的诱导和复苏

Induction and resuscitation of viable but non-culturable state of Staphylococcus aureus

摘要

摘要: 研究金黄色葡萄球菌活的非可培养状态（viable but non-culturable state,VBNC）的诱导和复苏方法。采用不同pH、温度、盐度、不同浓度的食品防腐剂山梨酸钾处理进行诱导;复苏则采用了直接升温处理、添加液体培养基升温处理、添加酵母液升温处理、添加吐温升温处理的方法。结果表明,经过不同温度和山梨酸钾诱导后的金黄色葡萄球菌进入了VBNC状态;其中诱导时间最短（76d）的条件是0.5mmol/L山梨酸钾、-20℃寡营养条件;采用BHI培养基升温的方法,以及添加0.5%和1%的吐温20和吐温80的培养液升温方法使进入VBNC状态的金黄色葡萄球菌得到复苏。VBNC状态的金黄色葡萄球菌在形态上发生了变化;复苏后的菌株除尿素酶阴性外,其他代谢特征均与标准菌株相同。

Abstract: The aim of this study was to investigate the VBNC state of Staphylococcus aureus and the resuscitation conditions.S.aureus was treated by different pH, salinity, temperature and different concentrations of preservative ( potassium sorbate) . The results showed that the strain was induced into the VBNC state under different concentrations of potassium sorbate and temperatures, and the minimum induction time was 76d at the condition of 0.5mmol /L potassium sorbate, - 20℃ and oligotrophy.The resuscitation of VBNC strain was success when being incubated in brain heart infusion ( BHI) medium at 37℃. Adding 0.5%, 1% Tween- 20 and Tween- 80 could help resuscitation.When the cells of S.aureus entered into the VBNC state, they all gradually changed from spherosome to irregular spherosome and decreased in size.The biochemical characteristics of the resuscitative cells were the same as the standard strain, except urease.