Recovery of Helicobacter pylori ATCC43504 from a viable but not culturable state: regrowth or resuscitation?
Recovery of Helicobacter pylori ATCC43504 from a viable but not culturable state: regrowth or resuscitation?
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DOI：
10.1111/j.1699-0463.1998.tb01386.x
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发表时间：
1998-05-01
期刊：
影响因子：
2.8
通讯作者：
Dainelli, B
中科院分区：
文献类型：
--
作者：
Cellini, L;Robuffo, I;Dainelli, B
对幽门螺杆菌球虫形态的形成和特征进行了研究。将幽门螺杆菌ATCC43504在加2%胎牛血清的布鲁氏菌肉汤中，在37℃、室温和4℃的微嗜气环境中培养，并于2、7、15、30和45天采集读数。在对照时期，进行了总计数和活菌计数、四氮唑盐活力试验和超微结构研究。在固体培养基上，幽门螺杆菌在室温和4℃孵育7天后变为不可培养，在37℃孵育15天后变为不可培养。在这些孵育时间，在相同条件下的液体培养基中继代培养后，在4℃孵育的培养基中检测到幽门螺杆菌的生长至第15天，在37℃和室温孵育的培养基中检测到生长至第30天。超微结构研究显示，细菌细胞的完整性逐渐降低，在30天和45天的孵育中保持稳定；30%的细菌全细胞在37℃和室温下孵育，50%的细菌在4℃下孵育。通过研究四氮唑盐INT（对碘硝基苯四氮唑紫）和CTC（氰基苯基四氮唑氯）还原成各自的甲酸盐来评估VNC（活的不可培养）状态的活力，这与细胞呼吸有关。在培养45 d时，当在固体或液体培养基中未观察到细菌再生时，采用不同的复苏方法来评估VNC幽门螺杆菌可能的复苏。在固体培养基上未观察到明显的生长。
Studies were conducted following the formation and characterization of the coccoid morphology of Helicobacter pylori. H. pylori ATCC43504 was incubated in brucella broth plus 2% fetal calf serum at three different temperatures: 37 degrees C, room temperature and 4 degrees C in a microaerophilic environment, and readings were taken at 2, 7, 15, 30 and 45 days. At control times, the total and the viable count, viability tests with tetrazolium salts, and ultrastructural studies were carried out. On solid media, H. pylori became nonculturable after 7 days of incubation at room temperature and 4 degrees C, and after 15 days of incubation at 37 degrees C. At these times of incubation, after subculturing in liquid medium under the same conditions, the growth of H. pylori was detected until the 15th day from cultures incubated at 4 degrees C and until the 30th day from cultures stored at 37 degrees C, and at room temperature. Ultrastructural studies showed a gradual reduction of integrity of bacterial cells that remained stable at 30 and 45 days of incubation: 30% of whole cells of bacteria incubated at 37 degrees C and room temperature and 50% in bacteria incubated at 4 degrees C. The viability of the VNC (viable nonculturable) state was assessed by studying the reduction of tetrazolium salts INT (p-iodonitrophenyl tetrazolium violet) and CTC (cyanoditolyl tetrazolium chloride) to their respective formazans and this was linked to the cellular respiration. At 45 days of incubation, when bacterial regrowth was not observed in solid or in liquid medium, different resuscitation methods were applied to evaluate a possible resuscitation of VNC H. pylori. No significant growth on solid medium was observed.