Wang Haili, Yan Yanfeng, Rong Dan, Wang Jing, Wang Hongduo, Liu Zizhong, Wang Jiaping, Yang Ruifu, Han Yanping
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China.
Department of Medical Monitoring and Support, Astronaut Center of China, Beijing, 100094, China.
Microbiologyopen. 2016 Oct;5(5):793-801. doi: 10.1002/mbo3.370. Epub 2016 May 16.
Biofilm formation is closely related to the pathogenetic processes of Klebsiella pneumoniae, which frequently causes infections in immunocompromised individuals. The immune system of astronauts is compromised in spaceflight. Accordingly, K. pneumoniae, which used to be isolated from orbiting spacecraft and astronauts, poses potential threats to the health of astronauts and mission security. Microgravity is a key environmental cue during spaceflight. Therefore, determining its effects on bacterial biofilm formation is necessary. In this study, K. pneumoniae ATCC BAA-1705 was exposed to a simulated microgravity (SMG) environment. K. pneumoniae grown under SMG formed thicker biofilms compared with those under normal gravity (NG) control after 2 weeks of subculture. Two indicative dyes (i.e., Congo red and calcofluor) specifically binding to cellulose fibers and/or fimbriae were utilized to reconfirm the enhanced biofilm formation ability of K. pneumoniae grown under SMG. Further analysis showed that the biofilms formed by SMG-treated K. pneumoniae were susceptible to cellulase digestion. Yeast cells mannose-resistant agglutination by K. pneumoniae type 3 fimbriae was more obvious in the SMG group, which suggests that cellulose production and type 3 fimbriae expression in K. pneumoniae were both enhanced under the SMG condition. Transcriptomic analysis showed that 171 genes belonging to 15 functional categories were dysregulated in this organism exposed to the SMG conditions compared with those in the NG group, where the genes responsible for the type 3 fimbriae (mrkABCDF) and its regulator (mrkH) were upregulated.
生物膜形成与肺炎克雷伯菌的致病过程密切相关,肺炎克雷伯菌常导致免疫功能低下个体感染。宇航员的免疫系统在太空飞行中会受到损害。因此,过去曾从轨道航天器和宇航员身上分离出的肺炎克雷伯菌,对宇航员的健康和任务安全构成潜在威胁。微重力是太空飞行期间的关键环境因素。因此,确定其对细菌生物膜形成的影响很有必要。在本研究中,将肺炎克雷伯菌ATCC BAA - 1705暴露于模拟微重力(SMG)环境中。传代培养2周后,与正常重力(NG)对照组相比,在SMG条件下生长的肺炎克雷伯菌形成了更厚的生物膜。使用两种特异性结合纤维素纤维和/或菌毛的指示染料(即刚果红和荧光增白剂)来再次确认在SMG条件下生长的肺炎克雷伯菌增强的生物膜形成能力。进一步分析表明,经SMG处理的肺炎克雷伯菌形成的生物膜易受纤维素酶消化。在SMG组中,肺炎克雷伯菌3型菌毛对酵母细胞的甘露糖抗性凝集更为明显,这表明在SMG条件下,肺炎克雷伯菌的纤维素产生和3型菌毛表达均增强。转录组分析表明,与NG组相比,暴露于SMG条件下的该生物体中属于15个功能类别的171个基因表达失调,其中负责3型菌毛(mrkABCDF)及其调节因子(mrkH)的基因上调。