Technical Abstract: We evaluated the effects of storage and handling conditions on the antimicrobial activity of biodegradable composite films (polylactic acid and sugar beet pulp) coated with allyl isothiocyanate (AIT). Polylactic acid (PLA) and chitosan were incorporated with AIT and coated on one side of the film. The films were treated at different storage conditions (storage time, storage temperature, and atmosphere) and handling conditions (washing, abrasion, and air blowing), and the antimicrobial activity of the films against Salmonella in tryptic soy broth was determined. The films (8.16 uL AIT/ cm2 surface area) significantly (p less than 0.05) inhibited the growth of Salmonella during 24 h incubation at 22C while the populations of Salmonella in controls increased from ca. 4 log CFU/ml to over 8 log CFU/ml, indicating a minimum 4 log CFU/ml reduction as compared with controls. Statistical analyses indicated that storage time, storage temperature, and surface abrasion affected antimicrobial activity of the films significantly (p less than 0.05). However, the difference in microbial reduction between these conditions was less than 0.5 log cycles. Results suggest that the films’ antimicrobial properties are stable under practical storage and handling conditions and these antimicrobial films have potential applications in food packaging.