Commercial producers of honey bee queens (Apis mellifera L.) have reported unexplained loss of immature queens during the larval or pupal stage. Many affected queen-rearing operations are situated among the almond orchards of California and report these losses in weeks after almond trees bloom. Almond flowers are a rich foraging resource for bees, but are often treated with fungicides, insecticides, and spray adjuvants during bloom. Anecdotal reports by queen producers associate problems in queen development with application of the fungicide Pristine (boscalid and pyraclostrobin) and spray adjuvants that are tank-mixed with it. To test the effect of these compounds on queen development, a new bioassay was developed in which queens are reared in closed swarm boxes for 4 d, until capping, with nurse bees fed exclusively on artificially contaminated pollen. Pollen was treated with four concentrations of formulated Pristine (0.4, 4, 40, and 400 ppm), a spray adjuvant (Break-Thru, 200 ppm), the combination of Pristine and spray adjuvant (400:200 ppm), the insect growth regulator insecticide diflubenzuron (100 ppm) as a positive control, or water as negative control. Chemical analysis revealed that low concentrations of pyraclostrobin (50 ppb), but no boscalid, were detectable in royal jelly secreted by nurse bees feeding on treated pollen. No significant difference in queen development or survival was observed between any of the experimental treatments and the negative control. Only diflubenzuron, the positive control, caused a substantial reduction in survival of immature queens.