Antibacterial electrospun fibrous membranes are widely applied as dressings for treatment the wounds and burns. However, developing long-term gelatin-based fibrous membrane is still the main challenge inhibiting their uses for long-term treatments. Novel antibacterial electrospun gelatin-based mats were introduced by combining gelatin, glycerol, glucose and silver nanoparticles (Ag NPs), which together could exhibit optimal physicochemical characteristics as long-term electrospun fibrous mats. Therefore, Ag NPs were synthesized in-situ within the acidic electrospun solutions during preparing electrospun gelatin-glycerol-Ag NPs (GEL-GLY-Ag) and gelatin-glycerol-glucose-Ag NPs (GEL-GLY-GLU-Ag) mats. Conventional spectroscopic techniques based on XRD, FTIR, SEM, DTA, and water uptake-degradation tests and antibacterial studies were used to characterize the preparations. The results showed that the electrospun gelatin-based composites mats revealed free beads dense fibrous textures, and exhibited a high water uptake and long-term degradation behavior. The Ag NPs could be successfully synthesized in-situ within electrospinning solutions and the results confirmed that the in-situ prepared Ag NPs enhanced the antibacterial activity of electrospun mats against positive and negative bacteria. (C) 2017 Elsevier B.V. All rights reserved.