Microscopic, electrochemical and surface characterization techniques were used to investigate the effects of iron reducing bacteria (IRB) biofilm on carbon steel corrosion for 72 and 168 h under batch conditions. The organic nutrient availability for the bacteria was varied to evaluate biofilms formed under nutritionally rich, as compared to nutritionally deficient, conditions. Focused ion beam-scanning electron microscopy (FIB-SEM) was used to investigate the effect of subsurface biofilm structures on the corrosion characteristics of carbon steel. Hydrated biofilms produced by IRB were observed under environmental scanning electron microscope (ESEM) with minimal surface preparation, and the elemental composition of the biofilms was investigated using energy dispersive spectroscopy (EDX). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) was used to provide information on the organic and inorganic chemical makeup of the biofilms. Electrochemical techniques employed for assessing corrosion, by open circuit potential, linear polarization and potentiodynamic polarization tests indicated no significant difference in the corrosion resistance for carbon steel in IRB-inoculated, compared to the abiotic solutions of common Postgate C after 72 and 168 h. However, the steel was found to be more susceptible to corrosion when the yeast extract was removed from the biotic environment for the 168 h test. In the absence of yeast nutrient, it is postulated that IRB received energy by transforming the protective film of Fe3+ into more soluble Fe2+ products.