Using Mössbauer resonance spectroscopy of 57Fe, we have determined the nature and distribution of major iron compounds in the magnetotactic bacterium Aquaspirillum magnetotacticum. In addition to magnetite (Fe3O4), cells contained a low-density hydrous ferric oxide, a high-density hydrous ferric oxide (ferrihydrite), and ferrous iron. Analysis at different temperatures of whole cells harvested early and late in growth, of mutant cells unable to synthesize magnetite, and of cell fractions enriched in 57Fe indicated that Fe3O4 precipitation resulted from partial reduction of the high-density hydrous ferric oxide precursor.