Induction and resuscitation of viable but nonculturable Salmonella enterica serovar typhimurium DT104
Induction and resuscitation of viable but nonculturable Salmonella enterica serovar typhimurium DT104
复制标题
DOI：
10.1128/aem.69.11.6669-6675.2003
复制
发表时间：
2003-11-01
影响因子：
4.4
通讯作者：
Joseph, SW
中科院分区：
文献类型：
--
作者：
Gupte, AR;de Rezende, CLE;Joseph, SW
检测了肠道沙门氏菌血清型鼠伤寒沙门氏菌DT 104 11601在最小化学成分确定的溶液中保持活力的能力。在不同离子浓度的微宇宙中，在不同的温度下，对生长和存活的定期监测表明，在5 ℃下，可培养的生物体逐渐减少（类似于235天）。保持在较高温度（21 ℃）下的微生物在接种后400天内显示出连续的、等效的CFU/ml（类似于10（6））。荧光显微镜与Baclight显示，non-cultureable细胞实际上是可行的，而与扫描电子显微镜观察显示，细胞保留了其结构的完整性。在胰蛋白胨大豆肉汤中对不可培养微生物（5 ℃）进行温度升高（56 ℃ +/- 0.5，15 s），然后立即接种到胰蛋白胨大豆琼脂（TSA）上，证明了复苏。有趣的是，S.在5 ℃（1 - 200天）或21 ℃（1 - 250天）下，来自微宇宙的肠道血清型鼠伤寒DT 104在间歇接种到添加过氧化氢酶的TSA上后没有显示出增强的生长。此外，细胞从21 degreesC微宇宙暴露于氧化和渗透胁迫表现出更大的抵抗压力，随着暴露时间的增加比最近生长的细胞。这种特殊菌株的特殊生存能力和恢复力可能部分反映了这种多重耐药生物体作为人类肠道疾病原因的重要性日益增加。S.肠道血清型鼠伤寒沙门氏菌DT 104 11601能够改变其生理特性，包括进入和恢复活的但不可培养的状态，这表明了沙门氏菌的总体可能性。肠道血清型鼠伤寒DT 104可能能够独特地响应于各种不利的环境条件。
Salmonella enterica serovar Typhimurium DT104 11601 was tested for its ability to maintain viability in minimal, chemically defined solutions. Periodic monitoring of growth and survival in microcosms of different ion concentrations, maintained at various temperatures, showed a gradual decline in culturable organisms (similar to235 days) at 5degreesC. Organisms maintained at a higher temperature (21degreesC) showed continuous, equivalent CFU per milliliter (similar to10(6)) up to 400 days after inoculation. Fluorescence microscopy with Baclight revealed that nonculturable cells were actually viable, while observations with scanning electron microscopy showed that the cells had retained their structural integrity. Temperature upshift (56degreesC +/- 0.5, 15 s) of the nonculturable organisms (5 degreesC) in Trypticase soy broth followed by immediate inoculation onto Trypticase soy agar (TSA) gave evidence of resuscitation. Interestingly, S. enterica serovar Typhimurium DT104 from the microcosms at either 5degreesC (1 to 200 days) or 21degreesC (1 to 250 days) did not show enhanced growth after intermittent inoculation onto catalase-supplemented TSA. Furthermore, cells from 21degreesC microcosms exposed to oxidative and osmotic stress showed greater resistance to stresses over increasing times of exposure than did recently grown cells. It is possible that the exceptional survivability and resilience of this particular strain may in part reflect the growing importance of this multidrug-resistant organism, in general, as a cause of intestinal disease in humans. The fact that S. enterica serovar Typhimurium DT104 11601 is capable of modifying its physiological characteristics, including entry into and recovery from the viable but nonculturable state, suggests the overall possibility that S. enterica serovar Typhimurium DT104 may be able to respond uniquely to various adverse environmental conditions.