Remediation measures for hexavalent chromium [Cr(VI)] are required for a safe environment. As a recent development in microbiology, bacterial biofilms are being studied as effective bioremediation agents. When bacteria are in fungal surface-attached biofilm mode, they are called fungal-bacterial biofilms (FBBs). They have not been tested for bioremediation so far. Hence, this study was conducted to develop FBBs and glass-wool-attached bacterial biofilms (BBs), and to evaluate Cr(VI) tolerability and removal of bacterial monocultures, BBs and FBBs. FBBs showed a significantly high level of Cr(VI) tolerance and resistance compared with its BBs or monocultures. After 10 days, up to 90% of Cr(VI) had been removed, which was significantly higher than that of BBs or its monocultures. Thus, it is clear that FBBs can be used as a novel tool to decontaminate Cr(VI) both in situ and ex situ.