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With the increasing demand for simple, efficient, environmentally friendly preparation methods to produce cellulose nanofibers for reinforcing a biodegradable film is increased, the role of nanofibers from the pure cellulose produced by bacteria becomes more important. This work characterized bacterial cellulose nanofibers disintegrated using a high shear homogenizer. These nanofibers, in 2.5, 5, and 7.5 mL suspensions, were mixed with PVA gel using ultrasonication. The resulting dried bionanocomposite film was also characterized. Adding nanofiber significantly increases (p ≤ 0.05) on tensile strength, thermal resistance, water vapor impermeability, and moisture resistance of PVA film but not strain at break. Tensile strength, tensile modulus, and elongation at the break of the 7.5 mL nanofiber reinforced film were 37.9 MPa (increased by 38%), 547.8 MPa (increased by 26%), and 10.7% (decreased from …