An anaerobic chamber to study the behaviour of microbes.

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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.