Due　to　the　large　specific　surface　area　and　rich　pore　structure,　chitosan　nanofiber　membrane　has　many　advantages　over　conventional　gel-like　or　film-like　products.　However,　the　poor　stability　in　acidic　solutions　and　relatively　weak　antibacterial　activity　against　Gram-negative　bacteria　severely　restrict　its　use　in　many　industries.　Here,　we　present　a　chitosan-urushiol　composite　nanofiber　membrane　prepared　by　electrospinning.　Chemical　and　morphology　characterization　revealed　that　the　formation　of　chitosan-urushiol　composite　involved　the　Schiff　base　reaction　between　catechol　and　amine　groups　and　the　self-polymerization　of　urushiol.　The　unique　crosslinked　structure　and　multiple　antibacterial　mechanisms　endowed　the　chitosan-urushiol　membrane　with　outstanding　acid　resistance　and　antibacterial　performance.　After　immersion　in　HCl　solution　at　pH　1,　the　membrane　main-tained　its　intact　appearance　and　satisfactory　mechanical　strength.　In　addition　to　its　good　antibacterial　perfor-mance　against　Gram-positive　Staphylococcus　aureus　(S.　aureus),　the　chitosan-urushiol　membrane　exhibited　synergistic　antibacterial　activity　against　Gram-negative　Escherichia　coli　(E.　coli)　that　far　exceeded　that　of　neat　chitosan　membrane　and　urushiol.　Moreover,　cytotoxicity　and　hemolysis　assays　revealed　that　the　composite　membrane　had　good　biocompatibility　similar　to　that　of　neat　chitosan.　In　short,　this　work　provides　a　convenient,　safe,　and　environmentally　friendly　method　to　simultaneously　enhance　the　acid　resistance　and　broad-spectrum　antibacterial　activity　of　chitosan　nanofiber　membranes.