The pathogenicity of enterohemorrhagic Escherichia coli O157:H7 were extensively studied by genomic and proteomic approaches. However, the possible virulence mechanism of E. coli O157:H7 in its hosts has never been studied using a metabolomic approach. In this study, the intracellular metabolites of C. elegans fed with pathogenic E. coli O157:H7 and non-pathogenic E. colt were profiled by gas chromatography/time-of-flight mass spectrometry. In C. elegans fed with O157:H7, the levels of metabolites related to the mammalian target of rapamycin (mTOR) signaling pathway, such as amino acids and glucose, highly increased. In addition, the levels of metabolites related to lipid oxidation and nucleotide salvage pathways increased. The metabolic intermediates of organic acidurias and atypical hemolytic uremic syndrome also increased when fed with O157:H7. However, the level of trehalose, an mTOR-independent autophagy enhancer, decreased in C. elegans fed with O157:H7. These results showed that infection with O157:H7 alters intracellular metabolite abundance in C. elegans. This study suggest that the metabolomics may be applied to elucidating the virulence mechanisms of pathogenic bacteria. (C) 2016 Elsevier Ltd. All rights reserved.