Aims: Studies were performed to demonstrate the efficacy of a novel electron beam irradiation system for the on-line sterilization of polymeric containers using energetic electrons directed through the container opening. Methods and Results: The distribution of dosage delivered during conveyance beneath the electron beam treatment system was determined for two sizes ( 8 or 16 ounces) of high-density polyethylene ( HDPE) blow-moulded bottles. The biological effects of treatment were then determined using Bacillus pumilus ATCC 27142 spores inoculated as 10 mul droplets dried ( i) onto the surface of flat coupons of the high-density polyethylene bottle material, and ( ii) onto the region of lowest delivered dose ( the bottom side wall) within each bottle. The inactivation obtained was determined by examining the level of survival after swab recovery of the inoculated spores, with reference to the level recovered from untreated control samples. Conclusions: The inactivation of B. pumilus spores during treatment was reproducible and proportional to the applied dose. In each instance, the logarithm of the surviving fraction of spores was well fitted by linear regression models and in good general agreement with values reported for inactivation of the same organism using gamma irradiation. Significance and Impact of the Study: The results of this work demonstrate the potential of a new physical method capable of high throughput for in-line sterilization of polymeric containers. The ability of the process to eliminate the washing step involved with traditional chemical sterilants, such as hydrogen peroxide, greatly reduces the size and environmental impact of these systems.