Excess unabsorbed iron in the gastrointestinal tract may select for enteric pathogens and increase the incidence and severity of infectious disease.Aspergillus oryzae(Ao) is a filamentous fungus that has the ability to accumulate and store large amounts of iron, and when used as a supplement or fortificant, has similar absorption to ferrous sulphate (FeSO4) in humans. The objective of this study was to determine the effect of iron-enrichedAo(Aoiron) compared with FeSO4on iron accumulation, growth and motility of the Gram-negative enteric pathogen,S.Typhimurium.S.Typhimurium was cultured in media containing no added iron or 1 μM elemental iron as eitherAoiron or FeSO4.S.Typhimurium cultured with FeSO4accumulated more iron than those cultured withAoiron. Genes regulated by the iron-activated transcriptional repressor, Fur, did not differ between control andAoiron, but decreased inS.Typhimurium cultured with FeSO4compared with both groups. Growth ofS.Typhimurium was greater when cultured with FeSO4compared withAoiron and control.S.Typhimurium swam faster, had greater acceleration and travelled further when cultured with FeSO4compared withAoiron and control; swim speed, acceleration and distance travelled did not differ betweenAoiron and control. These findings provide evidence thatAoiron reduces the virulence of a common enteric pathogenin vitro. Further research is required to determine whether iron-enrichedAois a suitable iron supplement to improve iron delivery in areas with a high infection burden.