Recovery in culture of viable but nonculturable Vibrio parahaemolyticus:: regrowth or resuscitation?
Recovery in culture of viable but nonculturable Vibrio parahaemolyticus:: regrowth or resuscitation?
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DOI：
10.1038/ismej.2007.1
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发表时间：
2007-06-01
期刊：
影响因子：
11
通讯作者：
Hervio-Heath, Dominique
中科院分区：
文献类型：
--
作者：
Coutard, Francois;Crassous, Philippe;Hervio-Heath, Dominique
本研究的目的是探索温度升高后可存活但不可培养 (VBNC) 副溶血性弧菌的可培养性恢复情况，并确定是否发生再生或复苏。副溶血性弧菌 Vp5 的临床菌株通过暴露于 4 摄氏度的人工海水 (ASW) 中而获得 VBNC。将 ASW 细胞悬浮液的等分试样（0.1、1 和 10 ml）置于 20 摄氏度和 37 摄氏度的高温下。监测等分试样中细胞在丰富培养基上集落形成的可培养性，并通过扫描 (SEM) 和透射 (SEM) 测量形态变化。 TEM）电子显微镜。 VBNC 细胞样本经过固定并通过 SEM 进行检查，揭示了由小细胞和较大扁平细胞组成的异质群体。温度升至 20 摄氏度或 37 摄氏度后 48 小时，使用 SEM 和 TEM 观察到细胞的二元裂变伸长和分裂，但仅限于 10 毫升等分试样。结果表明，部分 VBNC 细胞能够进行细胞分裂。结论是副溶血弧菌的一部分 VBNC 细胞在低温下仍能存活。温度升高后，这些细胞的重新生长，而不是初始接种物中所有细菌的复苏，似乎是副溶血性弧菌 VBNC 细胞可培养性恢复的原因。推测 VBNC 细胞滤液中的营养物质可以促进温度响应细胞的生长，细胞分裂通过二元裂变发生，但也包括非典型、不对称的细胞分裂。
The objective of this study was to explore the recovery of culturability of viable but nonculturable ( VBNC) Vibrio parahaemolyticus after temperature upshift and to determine whether regrowth or resuscitation occurred. A clinical strain of V. parahaemolyticus Vp5 was rendered VBNC by exposure to artificial seawater ( ASW) at 4 degrees C. Aliquots of the ASW suspension of cells ( 0.1, 1 and 10 ml) were subjected to increased temperatures of 20 degrees C and 37 degrees C. Culturability of the cells in the aliquots was monitored for colony formation on a rich medium and changes in morphology were measured by scanning ( SEM) and transmission ( TEM) electron microscopy. Samples of VBNC cells were fixed and examined by SEM, revealing a heterogeneous population comprising small cells and larger, flattened cells. Forty-eight hours after temperature upshift to 20 degrees C or 37 degrees C, both elongation and division by binary fission of the cells were observed, employing SEM and TEM, but only in the 10-ml aliquots. The results suggest that a portion of VBNC cells is able to undergo cell division. It is concluded that a portion of VBNC cells of V. parahaemolyticus subjected to cold temperatures remain viable. After temperature upshift, regrowth of those cells, rather than resuscitation of all bacteria of the initial inoculum, appears to be responsible for recovery of culturability of VBNC cells of V. parahaemolyticus. Nutrient in filtrates of VBNC cells is hypothesized to allow growth of the temperature-responsive cells, with cell division occurring via binary fission, but also including an atypical, asymmetric cell division.