An anaerobic chamber to study the behaviour of microbes.

结核病、腹泻和机会性感染等传染病是世界十大死亡原因之一。微生物还引起许多农作物和牲畜疾病，造成严重的经济损失，威胁到数百万人的食物摄入。抗生素耐药性的增加也是一个主要威胁，需要增加资源来研究细菌。依赖于荧光蛋白表达的基因编码报告对于理解微生物的生理是有帮助的。报告基因的发展可能会导致一些重要的应用，比如发现新的抗感染药物和理解基本的生物过程，比如细菌是如何在作物上定植、在食物上生长或感染人类的。然而，许多微生物生活在厌氧条件下，在这种条件下，经典的荧光蛋白是不活跃的。因此，我们必须找到替代这些传统方法的办法。参与该项目的研究小组正在分别研究与机会性肺部感染和腹泻疾病有关的铜绿假单胞菌和福氏志贺氏菌。我们的目标是研究它们在生命周期中遇到的厌氧条件下的行为。为此，我们要求获得厌氧室，以便在缺氧条件下进行实验。我们将使用这种基础设施来测试基于不需要氧气发光的新型荧光蛋白的基因报告。这些新的基因报告和其他方法将用于研究铜绿假单胞菌生物膜的形成和弗氏单胞菌3型分泌系统的功能。参与该项目的两个研究小组都隶属于渥太华大学，在微生物学方面拥有世界知名的专业知识。所要求的基础设施将与专门用于微生物活体成像的显微镜协同作用。每年将对大约15名高素质人员进行这方面的培训。不受限制地使用这些基础设施将使这些研究生能够毫不拖延地完成他们的培训。过去十年的发现表明，在植物、动物或环境中的微生物群中存在大量严格或兼性厌氧菌。这些认识刺激了许多学术或私人对厌氧菌的研究。在此基础设施上接受培训的个人获得的独特专业知识将使他们成为就业市场上备受追捧的资产。考虑到在其环境中暴露于厌氧条件下的微生物的绝对数量，我们期望这里开发的方法和报告将对研究其他微生物非常有用，这些微生物以以前不可能的方式威胁到公众健康和加拿大经济许多部门的财务可行性。

Infectious diseases, such as tuberculosis, diarrhea and opportunistic infections, are among the top-ten causes of death in the world. Microbes are also causing numerous crops and livestock diseases, which are responsible for severe economic losses and threaten the food intake of millions of people. The rise in antibiotic resistance is also a major threat that warrants increasing resources to study bacteria. Genetically encoded reporters relying on the expression of fluorescent proteins are instrumental to understand the physiology of microbes. Reporter development can lead down the road to important applications such as the discovery of novel anti-infective drugs and the understanding of fundamental biological processes such as how do bacteria colonize crop plants, grow on food or infect humans. Nonetheless, many microbes live in anaerobic conditions in which classical fluorescent proteins are inactive. We must therefore find alternatives to these conventional approaches. The research groups implicated in this project are studying bacteria Pseudomonas aeruginosa and Shigella flexneri implicated in opportunistic lung infections and diarrheal diseases, respectively. Our goal is to study their behavior in anaerobic conditions encountered during their lifecycle. To do this, we are requesting the acquisition of an anaerobic chamber, in order to perform experiments in anoxic conditions. We will use this infrastructure to test genetic reporters based on novel fluorescent proteins that do not require oxygen to emit light. These novel genetic reporters and other methods will be used to study the formation of biofilms by P. aeruginosa and the functioning of the Type 3 Secretion System of S. flexneri. Both research groups implicated on this project are affiliated to the University of Ottawa and possess world renown expertise in microbiology. The infrastructure requested will strongly synergize with a microscope dedicated to live imaging of microbes. Approximately 15 highly qualified personnel will be trained on this infrastructure every year. Unfettered access to this infrastructure will allow these graduate students to complete their training without delay. Discoveries in the last ten years have demonstrated the high abundance of strict or facultative anaerobic bacteria in the microbiota of plants, animals or in the environment. These realizations have spurred many academic or private efforts to study anaerobic bacteria. The unique expertise acquired by individuals trained on this infrastructure will make them highly sought after assets on the job market. Given the sheer number of microbes exposed to anaerobic conditions in their environment, we expect that the methods and reporters developed here will be extremely useful to study other microbes that threaten public health and the financial viability of many sectors of the Canadian economy in ways that were previously impossible.