Induction, resuscitation and quantitative real-time polymerase chain reaction analyses of viable but nonculturable Vibrio vulnificus in artificial sea water
Induction, resuscitation and quantitative real-time polymerase chain reaction analyses of viable but nonculturable Vibrio vulnificus in artificial sea water
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DOI：
10.1007/s11274-014-1640-1
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发表时间：
2014-08-01
影响因子：
4.1
通讯作者：
Bandekar, Jayant R.
中科院分区：
文献类型：
--
作者：
Rao, Namrata V.;Shashidhar, Ravindranath;Bandekar, Jayant R.
创伤弧菌是一种重要的食源性致病菌，在低温和低营养胁迫条件下可进入活的但不可培养（VBNC）状态。本研究考察了诱导创伤弧菌YJ 016复苏所需的VBNC状态和温度的诱导时间。还检查了VBNC状态期间和复苏细胞中细胞形态和基因表达的变化。在4A ℃人工海水中培养的创伤弧菌经过相当长的时间（70天）后进入VBNC状态。从VBNC诱导温度（4A ° C）增加6A ° C的温度导致VBNC细胞的复苏;然而，当VBNC细胞在23 A ° C保持24小时时观察到最大复苏。VBNC细胞形态由逗点状变为球粒状。用创伤弧菌细胞可以进行两轮VBNC诱导和复苏;然而，复苏的细胞数量逐渐减少，190天后细胞未能复苏。与膜蛋白[porinH]相关的基因显著上调（10.4倍），ompU（2.9倍）]，调节蛋白[envZ（5.6倍），toxR（4.5倍），toxS（4.8倍）]、氧化应激相关蛋白katG（2.3倍），细胞分裂/维持蛋白[ftsZ（4.3），MreB（6.5倍）]和复苏启动子因子yeaZ（四倍），表明这些基因在复苏期间发挥作用。基因表达的数据，这里提出的将加强我们的理解从VBNC状态的创伤弧菌复苏。研究结果也强调了在海鲜储存过程中保持低温的重要性。
Vibrio vulnificus, an important food-borne pathogen, is known to enter viable but nonculturable (VBNC) state under low temperature and low nutrition stress conditions. Present study examined the time required for induction of VBNC state and temperature which induces resuscitation of V. vulnificus YJ016. The change in cell morphology and gene expression during VBNC state and in resuscitated cells was also examined. V. vulnificus incubated in artificial sea water at 4 A degrees C entered VBNC state after considerably extended time (70 days). An increase in temperature by 6 A degrees C from the VBNC induction temperature (4 A degrees C) resulted in resuscitation of VBNC cells; however, maximum resuscitation was observed when VBNC cells were held at 23 A degrees C for 24 h. VBNC cells changed their morphology from comma shape to coccoid shape. Two rounds of induction of VBNC and resuscitation were possible with V. vulnificus cells; however, there was progressive reduction in number of resuscitated cells and after 190 days cells failed to resuscitate. Significant up-regulation of genes related to membrane proteins [porinH (10.4-fold), ompU (2.9-fold)], regulatory proteins [envZ (5.6-fold), toxR (4.5-fold), toxS (4.8-fold)], oxidative stress related protein katG (2.3-fold), cell division/maintenance proteins [ftsZ (4.3), mreB (6.5-fold)] and resuscitating promoter factor yeaZ (fourfold) was observed during resuscitation with respect to VBNC state indicating that these genes play a role during resuscitation. Gene expression data presented here would enhance our understanding of resuscitation of V. vulnificus from VBNC state. The results also highlight the importance of maintenance of low temperature during storage of seafood.