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首页> 外文期刊>Microbial Pathogenesis >A non-invasive in vivo imaging system to study dissemination of bioluminescent Yersinia pestis CO92 in a mouse model of pneumonic plague.
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A non-invasive in vivo imaging system to study dissemination of bioluminescent Yersinia pestis CO92 in a mouse model of pneumonic plague.

机译：一种非侵入性的体内成像系统，用于研究鼠疫性鼠疫小鼠模型中生物发光鼠疫耶尔森菌CO92的传播。

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The gold standard in microbiology for monitoring bacterial dissemination in infected animals has always been viable plate counts. This method, despite being quantitative, requires sacrificing the infected animals. Recently, however, an alternative method of in vivo imaging of bioluminescent bacteria (IVIBB) for monitoring microbial dissemination within the host has been employed. Yersinia pestis is a Gram-negative bacterium capable of causing bubonic, septicemic, and pneumonic plague. In this study, we compared the conventional counting of bacterial colony forming units (cfu) in the various infected tissues to IVIBB in monitoring Y. pestis dissemination in a mouse model of pneumonic plague. By using a transposon mutagenesis system harboring the luciferase (luc) gene, we screened approximately 4000 clones and obtained a fully virulent, luc-positive Y. pestis CO92 (Y. pestis-luc2) reporter strain in which transposition occurred within the largest pMT1 plasmid which possesses murine toxin and capsular antigen encoding genes. The aforementioned reporter strain and the wild-type CO92 exhibited similar growth curves, formed capsule based on immunofluorescence microscopy and flow cytometry, and had a similar LD50. Intranasal infection of mice with 15 LD50 of CO92-luc2 resulted in animal mortality by 72 h, and an increasing number of bioluminescent bacteria were observed in various mouse organs over a 24-72 h period when whole animals were imaged. However, following levofloxacin treatment (10 mg/kg/day) for 6 days 24 h post infection, no luminescence was observed after 72 h of infection, indicating that the tested antimicrobial killed bacteria preventing their detection in host peripheral tissues. Overall, we demonstrated that IVIBB is an effective and non-invasive way of monitoring bacterial dissemination in animals following pneumonic plague having strong correlation with cfu, and our reporter CO92-luc2 strain can be employed as a useful tool to monitor the efficacy of antimicrobial countermeasures in real time.
机译：用于监测感染动物中细菌传播的微生物学金标准一直是可行的板数。尽管是定量的，但是该方法需要牺牲被感染的动物。然而，最近，已经采用了生物发光细菌（IVIBB）的体内成像的替代方法，以监测宿主内的微生物传播。鼠疫耶尔森氏菌是一种革兰氏阴性细菌，能够引起鼠疫，败血症和肺炎鼠疫。在这项研究中，我们比较了在感染鼠疫的小鼠模型中，将各种感染组织中细菌菌落形成单位（cfu）的常规计数与IVIBB相比，以监测鼠疫耶尔森菌的传播。通过使用具有荧光素酶（luc）基因的转座子诱变系统，我们筛选了大约4000个克隆，并获得了完全有毒的，luc阳性的鼠疫耶尔森氏菌CO92（Y. pestis-luc2）报告基因菌株，其中转座发生在最大的pMT1质粒内它具有鼠毒素和荚膜抗原编码基因。上述报告菌株和野生型CO92表现出相似的生长曲线，基于免疫荧光显微镜和流式细胞术形成胶囊，并具有相似的LD 50 。用15 LD 50 的CO92-luc2鼻内感染可导致动物在72小时内死亡，整个动物在24-72小时内观察到各种小鼠器官中的生物发光细菌数量增加被成像。然而，左氧氟沙星在感染后24小时（10 mg / kg /天）治疗6天后，在感染72小时后未观察到发光，这表明所测试的抗菌素杀死细菌阻止了它们在宿主周围组织中的检测。总的来说，我们证明了IVIBB是监测与cfu密切相关的肺炎鼠疫后动物中细菌传播的有效且非侵入性的方法，我们的报道者CO92-luc2菌株可以用作监测抗菌对策功效的有用工具实时。

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