Antimicrobial packaging is a promising type of active packaging, which has an antimicrobial agent incorporated into the polymer matrix, capable to eliminate or inhibit deteriorating and/or pathogenic microorganisms. The zinc compounds are antimicrobial agents commonly used to confer this feature. The objective of this work was to evaluate the antimicrobial activity of biodegradable Ecoflex® films with 1% and 2% (w/w) zinc oxide nanoparticles (ZnO NPs) and 1% and 2% (w/w) glass microparticles doped with ionic zinc (Zn MCs), against Staphylococcus aureus and Escherichia coli. The sample’s biodegradation in soil also were studied. Microbiological results showed that biodegradable polymeric films obtained were effective to inhibit and/or kill S. aureus and E. coli at the initial concentration of 104 CFU.mL-1 after 24 h of contact. When evaluated the biodegradation of the polymers containing the additives, the results showed that the addition of zinc compounds to the biodegradable polymer did not affect its biodegradability characteristic. The antimicrobial biodegradable films developed have potential application to the food packaging industry.