This study systematically evaluated the effect of ferric iron on sulfate reduction to sulfide, feed digestion and fermentation, methane production, and populations of select ruminal microbes using in vitro rumen cultures. Ferric oxide (Fe2O3) and ferric citrate (C6H5FeO7) at six concentrations (0, 25, 50, 100, 150, and 200 mg/L as Fe3+) were tested. Ferric iron decreased production of both H2S gas in culture headspace (up to 71.9%) and aqueous sulfide (up to 80.8%), without adversely affecting other fermentation parameters, with ferric citrate being more effective than ferric oxide. Total archaeal population was increased by ferric citrate, but methane production was not affected significantly. The population of sulfate reducing bacteria was affected differently by ferric oxide than by ferric citrate. The results of this study could guide future in vivo studies to develop effective solutions to abate sulfur-associated polioencephalomalacia in cattle fed high-sulfur diet such as dried distiller's grains with solubles. (C) 2016 Elsevier Ltd. All rights reserved.