Microfibers (MFs) are widely existed in indoor air; however, characteristic of microbiota on MFs is largely unknown. In this study, air-borne MFs were collected from fallout or air-conditioner (AC) filter dust in three types of indoor space including living room, dormitory and office. Both plastic and natural MFs were identified by Fourier transform infrared spectroscopy. Ultramicroscopic observation showed dense biofilms adhering on surfaces of MFs. Fallout MFs contained more bacteria but fewer fungi than MFs from AC filter dust. MFs-loaded bacteria were of highest abundance in living rooms, following dormitories and offices. Bacterial community and its diversity were further analyzed by 16S rRNA High-throughput sequencing. Up to 4540 of bacterium OTUs were shared in these MFs samples, unique OTUs in fallout and AC filter samples accounting for 26.3 % and 25.7 % of the total. Compared to MFs fallout, AC filter MFs contained more species of pathogenic bacteria, such as Betaproteobacteriales and Ralstonia, with obviously different β-diversity between two groups. Phenotypic analysis showed that fallout and AC filter MFs bacteria presented high index values of film formation, oxidative stress tolerance and potential pathogenicity. Overall, these results suggest that abundant bacteria including pathogen can be loaded on MFs, and would pose health risks through delivery of indoor MFs.