In this work, zinc oxide (ZnO) micron and nano sized-particles with different morphologies were synthesized by aqueous precipitation and evaluated as antimicrobial agents against foodborne pathogens. The most effective bactericide system was selected to prepare active poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) films by three different methods (i) direct melt-mixing, (ii) melt-mixing of preincorporated ZnO into PHBV18 (18 mol% valerate content) fiber mats made by electrospinning, and, (iii) as a coating of the annealed electrospun PHBV18/ZnO fiber mats over compression molded PHBV. Results showed that ZnO successfully improved the thermal stability of the PHBV18, being the preincorporation method the most efficient in mitigating the negative impact that the PHBV18 had on the thermal stability, barrier and optical properties of the PHBV films. Similar behavior was found for the coating structure although this film showed effective and prolonged antibacterial activity against Listeria monocytogenes. This study highlights the suitability of the PHBV/ZnO nanostructures for active food packaging and food contact surface applications. (C) 2016 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.