AbstractVinegar flies (Diptera: Drosophilidae) are well known to be associated with yeasts, which provide important nutrients and emit attractive semiochemicals. Drosophila suzukii (Matsumura) has become a major pest of berries and cherries around the world, requiring intensive management to maintain fruit quality. Although insecticides remain a dominant control approach, disruption of fly–yeast–host interactions remains a promising avenue for reducing the economic impact of this pest. We conducted field and laboratory experiments to explore whether a crop sterilant (peroxyacetic acid and hydrogen peroxide) developed for disease control can affect D. suzukii. In 2 yr of field tests in highbush blueberries, we found significantly lower infestation by D. suzukii in plots treated with the crop sterilant, both alone and in a rotation program with zeta-cypermethrin. When shoots from treated plots were tested in no-choice bioassays, crop sterilant treatments did not affect adult mortality or oviposition, but they reduced infestation. To explore the mechanisms in the laboratory, we found that the crop sterilant did not affect adult mortality, nor oviposition on treated fruit under no-choice settings, but adult flies settled and oviposited less on treated fruit in choice settings. When the crop sterilant was applied to colonies of Hanseniaspora uvarum (Niehaus) (Saccharomycetales: Saccharomycodaceae) and Issatchenkia terricola (Van der Walt) (Saccharomycetales: Saccharomycetacea) yeasts that are attractive and provide nutrition to D. suzukii, there was a dose-dependent inhibition of their growth. We highlight the potential for microbial management as a component of integrated pest management programs and prioritize research needs to incorporate this approach into control programs.