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Microarray analysis of  Pseudomonas aeruginosa  mRNA transcripts expressed  in vivo  during animal infection has not been previously used to investigate potential virulence factors needed in this setting. We compared mRNA expression in bacterial cells recovered from the gastrointestinal (GI) tracts of  P. aeruginosa -colonized mice to that of  P. aeruginosa  in the drinking water used to colonize the mice. Genes associated with biofilm formation and type III secretion (T3SS) had markedly increased expression in the GI tract. A non-redundant transposon library in  P. aeruginosa  strain PA14 was used to test mutants in genes identified as having increased transcription during  in vivo  colonization. All of the Tn-library mutants in biofilm-associated genes had an attenuated ability to form biofilms  in vitro , but there were no significant differences in GI colonization and dissemination between these mutants and WT  P. aeruginosa  PA14. To evaluate T3SS factors, we tested GI colonization and neutropenia-induced dissemination of both deletional (PAO1 and PAK) and insertional (PA14) mutants in four genes in the  P. aeruginosa  T3SS,  exoS  or  exoU ,  exoT , and  popB . There were no significant differences in GI colonization among these mutant strains and their WT counterparts, whereas rates of survival following dissemination were significantly decreased in mice infected by the T3SS mutant strains. However, there was a variable, strain-dependent effect on overall survival between parental and T3SS mutants. Thus, increased transcription of genes during  in vivo  murine GI colonization is not predictive of an essential role for the gene product in either colonization or overall survival following induction of neutropenia.

Utility of In Vivo Transcription Profiling for Identifying Pseudomonas aeruginosa Genes Needed for Gastrointestinal Colonization and Dissemination