Understanding pathogen transmission could illuminate new methods for disease prevention. A case in point is chalkbrood in the alfalfa leafcutting bee [Megachile rotundata (F.)]. Propagation of this solitary bee is severely hampered by chalkbrood, a larval disease caused by Ascosphaera aggregata (Ascomycota). Alfalfa leafcutting bees nest in existing cavities in wood or hollow reeds and overwinter as larvae. In the early summer, emerging adults frequently must chew through dead, diseased siblings that block their exit, becoming contaminated with chalkbrood spores in the process. When alfalfa leafcutting bees are used as a commercial pollinator, the cocoons are removed from nesting boards to reduce chalkbrood transmission, but the disease is still common. To determine if these removed cocoons (called loose cells) are an important source of disease transmission, they were disinfected with a fungicide before bees were incubated, and released in the field. Chalkbrood prevalence among the progeny of the treated bees was reduced up to 50% in one field trial, but not significantly when tested in an on-farm trial. Thus, substantial disease transmission still occurred when the loose cells were disinfected, and even when clean nesting materials were used. In conclusion, pathogen transmission must still be occurring from another source that has yet to be identified. Another possible source of transmission could arise from bees that emerge midsummer in populations with a high percent of multivoltinism, but dirty nesting boards and feral bees also may be minor sources of transmission.