Dietary antioxidants may have a role in preventing some of the chronic diseases in humans resulting from free radical oxidation of lipids and other cellular components. Blueberries (Vaccinium L. sp.) are considered one of the best fresh fruit sources of antioxidants, and there is the potential to increase the antioxidant activity further through breeding. Thus, the variability of fruit antioxidant activity (AA) was examined among a set of 16 highbush and interspecific hybrid cultivars grown at locations in Minnesota (MN), Michigan (MI), and Oregon (OR) over 2 years (1998 and 1999) to determine effects of genotype, year, and location. Nine cultivars were common to all three locations in both years. Antioxidant activity, total phenolic content (TPH), and total anthocyanin content (ACY), were determined in triplicate samples from each genotype. Cultivars differed significantly (P ≤ 0.05) in AA, TPH, and ACY both within and over locations. The single location mean AA for all cultivars changed significantly between the 2 years in OR and in MI, while the single location mean for TPH differed between the 2 years in MN and MI. Changes in cultivar rank were significant for AA, TPH, and ACY between years within each location. Significant changes in rank for TPH and ACY were also noted between pairs of locations as well. Pearson's correlation for AA (based on cultivar means) appeared highest between MN and OR (r = 0.90) and MN and MI (r = 0.69) in 1998; correlations between locations for the combined years were 0.74 for MN and OR, 0.55 for MN and MI and 0.45 for MI and OR. For the group of nine cultivars, AA correlated well with TPH within each location, with r ranging from 0.67 to 0.95 for data from individual and combined years. Correlation of AA with ACY at each location was lower than that for AA with TPH, in both individual and combined years. This study demonstrates significant genotype x environment interaction for AA in blueberry.