Purpose: The objective of the present investigation was to evaluate the antibacterial properties and the biocompatibility of composite electrospun nanofibrous membranes (NFMs) with -low-molecular-weight fish scale collagen peptides (FSCP) and chito-oligosaccharide (COS), to determine their potential for use as wound dressings. Methods: Low-molecular-weight FSCP were combined with COS to prepare nanofibers by electrospinning, and polyvinyl alcohol (PVA) was used for enhancing fiber-forming ability. Transmission electron microscope and scanning electron microscope methods were used to observe bacterial adhesion and the bacterial cell membrane. Fibroblast cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results: The best FSCP/COS mass ratio for electrospinning was 2: 1, and the nanofibers had small dimensions ranging from 50 to 100 nm. The NFM showed good antibacterial activities against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli. The antibacterial activity against S. aureus was higher than against E. coli. The pili and adhesive fimbriae of E. coli promoted bacterial adhesion to the NFM surfaces, and S. aureus biofilms aided S. aureus adhesion on the surface of NFMs. Damage to the bacterial cell membrane indicates that the NFMs could lead to the release of intracellular materials, particularly with S. aureus. In addition, FSCP/COS NFM rapidly increased the permeability of the outer membranes of E. coli. The electrospun NFM with FSCP and COS had good biocompatibility in vitro and supported proliferation of human skin fibroblasts. Conclusion: FSCP are superior to mammalian collagen, and have feasibility and potency for wound dressings. FSCP/COS NFMs had good anti-bactericidal activity that improved with increased COS, and showed good biocompatibility in vitro and supported the proliferation of fibroblasts.