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Determination of spatial and temporal colonization of enteropathogenic E. coli and enterohemorrhagic E. coli in mice using bioluminescent in vivo imaging

机译:使用生物发光体内成像法测定小鼠肠道致病性大肠杆菌和肠出血性大肠杆菌的时空定殖

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摘要

Infectious diarrhea is a major contributor of child morbidity and mortality in developing nations. Murine models to study the pathogenesis of infectious diarrhea caused by organisms such as enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC) are not fully characterized. More emphasis has been placed on infection of mice with the murine specific pathogen Citrobacter rodentium. While these three organisms are genetically related they are not identical. Our goal was to better characterize the murine model of EPEC and EHEC infection by using bioluminescent bacteria to determine temporal and spatial colonization of these two human pathogens. EPEC and EHEC were transformed with a bacterial luciferase expression plasmid containing the constitutive OmpC promoter. C57BL/6 mice were orally inoculated with bioluminescent EPEC or EHEC and bacterial localization in the intestine was monitored ex vivo and in vivo by IVIS. At 3 days after infection, EPEC, EHEC and Citrobacter rodentium were all localized in the cecum and colon. EPEC colonization peaked at day 2–3 and was undetectable by day 7. The bioluminescent EPEC adheres to the cecum and colon of the mouse intestine. However, when EPEC infected mice were administered xylazine/ketamine for in vivo live imaging, the EPEC persisted at high densities for up to 31 days. This is the first report of a bioluminescent imaging of luciferase expressing EPEC in a mouse model.
机译:传染性腹泻是发展中国家儿童发病率和死亡率的主要诱因。鼠模型研究由诸如肠道致病性大肠杆菌(EPEC)和肠出血性大肠杆菌(EHEC)之类的生物引起的感染性腹泻的发病机理尚不完整。已经将更多的重点放在用鼠特异性病原体啮齿类柠檬酸杆菌感染小鼠上。虽然这三种生物是遗传相关的,但它们并不相同。我们的目标是通过使用生物发光细菌确定这两种人类病原体的时间和空间定殖,更好地表征EPEC和EHEC感染的小鼠模型。用含有组成型OmpC启动子的细菌荧光素酶表达质粒转化EPEC和EHEC。用生物发光EPEC或EHEC口服接种C57BL / 6小鼠,并通过IVIS进行离体和体内监测肠道中细菌的定位。感染后第3天,EPEC,EHEC和啮齿类柠檬酸杆菌都位于盲肠和结肠。 EPEC的定植在第2-3天达到峰值,到第7天则无法检测到。生物发光的EPEC附着在小鼠肠的盲肠和结肠上。但是,当EPEC感染的小鼠接受甲苯噻嗪/氯胺酮进行体内活体成像时,EPEC可以高密度持续长达31天。这是在小鼠模型中表达荧光素酶的荧光素酶的生物发光成像的首次报道。

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