Loukil Ahmed, Lalaoui Rym, Bogreau Hervé, Regoui Sofiane, Drancourt Michel, Hammoudi Nassim
Aix Marseille Univ., IRD, APHM, MEPHI, Marseille, France.
IHU-Méditerranée Infection, Marseille, France.
Am J Trop Med Hyg. 2022 Apr 11;107(1):89-93. doi: 10.4269/ajtmh.21-1327.
Whether Mycobacterium ulcerans, the etiological agent of Buruli ulcer in numerous tropical countries, would exist in a dormant state as reported for closely related Mycobacterium species, has not been established. Six M. ulcerans strains were exposed to a progressive depletion in oxygen for 2 months, using the Wayne model of dormancy previously described for M. tuberculosis, and further examined by microscopy after staining of dynamic, dormant, and dead mycobacteria (DDD staining), microcalorimetry and subculture in the presence of dead and replicative M. ulcerans as controls. Mycobacterium ulcerans CU001 strain died during the progressive oxygen depletion and four of five remaining strains exhibited Nile red-stained intracellular lipid droplets and a 14- to 20-day regrowth when exposed to ambient air, consistent with dormancy. A fifth M. ulcerans 19423 strain stained negative in DDD staining and slowly regrew in 27 days. Three tested M. ulcerans strains yielded microcalorimetric pattern similar to that of the negative (dead) homologous controls, differing from that of the homologous positive (replicative) controls. The relevance of these experimental observations, suggesting a previously unreported dormancy state of M. ulcerans, warrants further investigations in the natural ecological niches where M. ulcerans thrive as well as in Buruli ulcer lesions.
在许多热带国家引发布氏溃疡的溃疡分枝杆菌是否会像与其亲缘关系密切的分枝杆菌物种那样处于休眠状态,目前尚未确定。利用先前针对结核分枝杆菌描述的韦恩休眠模型,将6株溃疡分枝杆菌菌株置于氧气逐渐耗尽的环境中2个月,然后在对活的、休眠的和死亡的分枝杆菌进行染色(DDD染色)后通过显微镜检查、微量热法以及在存在死亡和增殖的溃疡分枝杆菌作为对照的情况下进行传代培养进一步检测。溃疡分枝杆菌CU001菌株在氧气逐渐耗尽过程中死亡,其余5株中的4株在暴露于环境空气时表现出尼罗红染色的细胞内脂滴以及14至20天的再生长,这与休眠一致。第五株溃疡分枝杆菌19423菌株在DDD染色中呈阴性,并在27天内缓慢再生长。3株受试溃疡分枝杆菌菌株产生的微量热图谱与阴性(死亡)同源对照相似,与同源阳性(增殖)对照不同。这些实验观察结果表明溃疡分枝杆菌存在此前未报道的休眠状态,这一现象的相关性值得在溃疡分枝杆菌大量繁殖的自然生态位以及布氏溃疡病变中进一步研究。