With the growing popularity of blueberries and the associated increase in blueberry imports and exports worldwide, delivering fruit with high quality, longer shelf-life, and meeting phytosanitary requirements has become increasingly important. The objective of this study was to determine the effects of electron beam irradiation using a new Electronic Cold-PasteurizationTM (ECPTM) technology on fruit quality, microbial safety, and postharvest disease development in two southern highbush blueberry cultivars, ‘Farthing’ and ‘Rebel’. Fruit packed in clamshells were subjected to four levels of ECPTM irradiation (0, 0.15, 0.5, and 1.0 kGy) and evaluated for fruit quality attributes, surface microbial load, and postharvest disease incidence during various storage times after treatment and cold storage. Overall, there was no effect of irradiation on visual fruit quality in either cultivar. Fruit firmness and skin toughness in ‘Farthing’ was reduced following irradiation at 1.0 kGy, but no such effect was observed in ‘Rebel’. Other fruit quality characteristics such as fruit weight, total soluble solids content, or titratable acidity were not affected. Irradiation at 1.0 kGy significantly reduced total aerobic bacteria and yeast on the fruit surface, and in the case of ‘Rebel’, also levels of total coliform bacteria. There was no significant effect of irradiation on postharvest disease incidence in these trials. Overall, data from this study suggests that an irradiation dose lower than 1.0 kGy using ECPTM can be useful for phytosanitary treatment in blueberry fruit while avoiding undesirable effects on fruit quality in a cultivar-dependent manner.