Abstract:[Background] Nano-particles produced by microorganisms in situ can enhance oil recovery. [Objective] To study the effects of iron nano-particles produced by microorganisms in situ on oil recovery by combining microbial production of nanoparticles with the technology of nano-fluids enhancing oil recovery. [Methods] We isolated the strains from shale rock debris and identified the strains by genome sequencing. Subsequently, we established the standard curve of Fe(II) concentration by the o-phenanthroline method to determine the Fe(III) reduction ability of the strain, and characterized the morphology and particle size of nano-particles produced by strain interaction with hematite (Fe2O3) by scanning electron microscopy, transmission electron microscopy, and Zeta potential analysis. Finally, we investigated the ability of the nano-particles to enhance oil recovery by determining the dynamic wetting angle at oil/water/rock interface and conducting oil droplet stripping and transport experiments. [Results] The isolated strain was identified as Shewanella chilikensis FR1. FR1 could rapidly reduce Fe2O3, with the iron reduction rate reaching 65.26%. After the strain was interacted with Fe2O3 for 10 days, a large number of spherical nano-particles were observed on the surface of hematite on the bacterial cell walls, inside of the cells, and eaten by bacteria. The particles with the size >1 000 nm accounted for 88.05% in the blank control without strain inoculation, and those in the bacterial inoculation group mainly had the size of 10–1 000 nm. The nanoparticles in the oil-rock interface could obviously change the wettability of oil-wetted interfaces and effectively strip crude oil on the solid interface. The nano-particles generated by FR1 in situ could be used for core flooding, decreasing the surface tension of the aqueous phase of the discharged fluid to 27.9–28.6 mN/m and increasing the oil recovery to 16.38%–17.76%. [Conclusion] The results of this study provide a new technical idea for microbial enhanced oil recovery (MEOR).