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The average density observed was only 3*10 4 CFU/g, which is several orders of magnitude below the critical level (~ 1*10 9 CFU/g ( Adding to these physiological barriers, whole-genome sequencing later revealed a major reason for this failure: VNP20009 contained unintentional spontaneous mutations in the rpoS gene, encoding the stress-response sigma factor, and a silent mutation in suhB , reducing its metabolic fitness as well as its ability to colonize hepatocytes (79)

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