The dissimilatory metal-reducing bacterium,Shewanella oneidensisMR-1 produced γ-MnS (rambergite) nanoparticles during the concurrent reduction of MnO2 and thiosulfate coupled to H2 oxidation. To investigate effect of direct microbial reduction of MnO2 on MnS formation, two MR-1 mutants defective in outer membranec-type cytochromes (ΔmtrC/ΔomcAand ΔmtrC/ΔomcA/ΔmtrF) were also used and it was determined that direct reduction of MnO2 was dominant relative to chemical reduction by biogenic sulfide generated from thiosulfate reduction. Although bicarbonate was excluded from the medium, incubations of strain MR-1 with lactate as the electron donor produced MnCO3 (rhodochrosite) as well as MnS in nearly equivalent amounts as estimated by micro X-ray diffraction (micro-XRD) analysis. It was concluded that carbonate released from lactate metabolism promoted MnCO3 formation and that Mn(II) mineralogy was strongly affected by carbonate ions even in the presence of abundant sulfide and weakly alkaline conditions expected to favour the precipitation of MnS. Formation of MnS, as determined by a combination of micro-XRD, transmission electron microscopy, energy dispersive X-ray spectroscopy, and selected area electron diffraction analyses was consistent with equilibrium speciation modelling predictions. Biogenic manganese sulfide may be a manganese sink in the Mn biogeochemical cycle in select environments such as deep anoxic marine basins within the Baltic Sea.