Miranda Regina L, Conway Tyrrell, Leatham Mary P, Chang Dong Eun, Norris Wendy E, Allen James H, Stevenson Sarah J, Laux David C, Cohen Paul S
Department of Cell and Molecular Biology, University of Rhode Island, Kingston, Rhode Island 02881, USA.
Infect Immun. 2004 Mar;72(3):1666-76. doi: 10.1128/IAI.72.3.1666-1676.2004.
Escherichia coli EDL933, an O157:H7 strain, is known to colonize the streptomycin-treated CD-1 mouse intestine by growing in intestinal mucus (E. A. Wadolkowski, J. A. Burris, and A. D. O'Brien, Infect. Immun. 58:2438-2445, 1990), but what nutrients and metabolic pathways are employed during colonization has not been determined. In this study, when the wild-type EDL933 strain was fed to mice along with an EDL933 DeltappsA DeltapckA mutant, which is unable to utilize tricarboxylic acid cycle intermediates and gluconeogenic substrates for growth, both strains colonized the mouse intestine equally well. Therefore, EDL933 utilizes a glycolytic substrate(s) for both initial growth and maintenance when it is the only E. coli strain fed to the mice. However, in the presence of large numbers of MG1655, a K-12 strain, it is shown that EDL933 utilizes a glycolytic substrate(s) for initial growth in the mouse intestine but appears to utilize both glycolytic and gluconeogenic substrates in an attempt to maintain colonization. It is further shown that MG1655 predominantly utilizes glycolytic substrates for growth in the mouse intestine whether growing in the presence or absence of large numbers of EDL933. Data are presented showing that although small numbers of EDL933 grow to large numbers in the intestine in the presence of large numbers of MG1655 when both strains are fed to mice simultaneously, precolonization with MG1655 affords protection against subsequent colonization by EDL933. Moreover, in mice that are precolonized with EDL933, small numbers of MG1655 are able to grow rapidly in the intestine and EDL933 is eliminated. In situ hybridization experiments using E. coli-specific rRNA probes showed that while MG1655 is found only in mucus, EDL933 is found both in mucus and closely associated with intestinal epithelial cells. The data are discussed with respect to competition for nutrients and to the protection that some intestinal commensal E. coli strains might afford against infection by O157:H7 strains.
大肠杆菌EDL933是一种O157:H7菌株,已知它通过在肠道黏液中生长而在经链霉素处理的CD-1小鼠肠道中定殖(E. A. 瓦多科夫斯基、J. A. 伯里斯和A. D. 奥布赖恩,《感染与免疫》58:2438 - 2445,1990),但在定殖过程中利用了哪些营养物质和代谢途径尚未确定。在本研究中,当将野生型EDL933菌株与无法利用三羧酸循环中间体和糖异生底物进行生长的EDL933 ΔppsA ΔpckA突变体一起投喂给小鼠时,两种菌株在小鼠肠道中的定殖情况同样良好。因此,当EDL933是唯一投喂给小鼠的大肠杆菌菌株时,它利用糖酵解底物进行初始生长和维持生长。然而,在存在大量K - 12菌株MG1655的情况下,结果表明EDL933利用糖酵解底物在小鼠肠道中进行初始生长,但似乎利用糖酵解和糖异生底物来维持定殖。进一步表明,无论是否在大量EDL933存在的情况下生长,MG1655在小鼠肠道中生长时主要利用糖酵解底物。所呈现的数据表明,当两种菌株同时投喂给小鼠时,尽管在大量MG1655存在的情况下少量EDL933在肠道中可大量生长,但预先用MG1655定殖可提供针对随后EDL933定殖的保护。此外,在预先用EDL933定殖的小鼠中,少量MG1655能够在肠道中快速生长,而EDL933被清除。使用大肠杆菌特异性rRNA探针进行的原位杂交实验表明,虽然MG1655仅存在于黏液中,但EDL933既存在于黏液中,也与肠道上皮细胞紧密相关。针对营养物质竞争以及一些肠道共生大肠杆菌菌株可能提供的针对O157:H7菌株感染的保护进行了讨论。