Two chitosan derivatives, namely, thiol-chitosan (TCs) and chitosan iodoacetamide (CsIA) were newly synthesized by reacting Cs with thiglycolic acid (TGA) and iodoacetic acid (IA) respectively. After being crosslinked with glutraldehyde (GA), the two derivatives were submitted to FT-IR and H1 NMR analysis for identification and characterization of their chemical features. As TCs and CsIA are water soluble, their electrospun nanofibres mats from aqueous solutions could be crosslinked and achieved using polyvinyl alcohol/Chitosan blend (PVA/Cs) polymers. Morphological structures of the obtained nanofibres and their webs were studied via those of TCs and CsIA free systems. The data also indicate that the crosslinked PVA/Cs/CsIA is more thermally stable than the crosslinked PVA/Cs/TCs and crosslinked PVA/Cs respectively. It was proved that the electrospun fibers containing TCs or CsIA display a superior antibacterial activity against negative bacteria E. Coli with a minimum inhibitory concentration (MIC) of 400μg/ml. These effects are rather in confirmation with bacterial kinetics essays which were also carried out in current work. Of particular interest is that the antimicrobial properties of fibers containing small concentration of either TCs or CsIA are much superior than those obtained with neat Cs electrospun nanofibres used as reference. By and large the results advocate the fibers webs containing TCs or CsIA as excellent candidates for wound dressing applications.