Nanofibrous films and coatings were fabricated by electrospinning and centrifugal spinning of aqueous solutions of poly(vinyl alcohol-co-vinyl acetate)/octadecyl amine-montmorillonite layered silicate nanocomposites with/without in situ generated silver nanoparticles (AgNPs). Characterization of nanofibers were realized by FTIR, XRD, SEM, and thermal analysis methods. It was found that AgNPs significantly accelerated phase separation process providing fine distribution of nanofibers. Antifungal and antimicrobial activities of nanocomposites investigated by using Candida Spp fungals, G-positive and G-negative microorganisms strongly depended on chemical/physical structural factors, and loading silver species. Observed effective properties of nanomaterials can be employed for many applications (e.g., air filtration, food packaging, biomedical).