Transgenic crops with plant-incorporated protectants are often more specific than synthetic insecticides and have the potential to reduce impacts on non-target organisms. In this study we assessed the impact of Cry3Aa and Cry3Bb1 coleopteran-active d-endotoxins on the bulb mite, Rhizoglypus robini. The effect of Cry3Aa prototoxin in solutions of the biopesticide Novodor® on mite survival was assessed in laboratory studies. Survival of R. robini exposed to Cry3Aa in a short-term contact and ingestion experiment was not affected, although R. robini was significantly affected by the insecticide Fipronil® used as a positive control. Similarly, R. robini exposed in a longer duration feeding trial to the Cry3Aa toxin in artificial diet were also not significantly affected. When Cry3Aa was tested on the positive control insect, Leptinotarsa decemlineata, reduced weight of larvae and increased mortality was recorded. The effect of Cry3Bb1 toxin in transgenic corn tissues on R. robini food choice was assessed in a laboratory study. In no-choice tests a greater proportion of R. robini were found on garlic roots than on Cry3Bb1 transgenic corn and a near-isogenic non-transgenic corn. In a choice test, more R. robini was recovered on garlic roots than on either corn variety, and on Cry3Bb1 corn than on non-transgenic corn. In large field plots using specific mite traps across the growing season, R. robini mite populations were not significantly different between Cry3Bb1 corn and non-transgenic corn alone or non-transgenic corn treated with different combinations of two insecticides. Our results, combined with results from other studies, suggest that transgenic plants expressing the Cry3Aa or Cry3Bb1 Bacillus thuringiensis toxins are likely to have negligible impact on R. robini mite populations.