Abstract Female multiple mating with different males (polyandry) can be advantageous because the resulting genetic heterogeneity among offspring reduces the effects of parasitism. However, the underlying assumption that offspring fathered by different males vary in their susceptibility to parasites is so far only supported indirectly. Here we tested this crucial assumption using data from a study on the bumblebee Bombus terrestris L. with queens inseminated with sperm of either one or several males that originated from different sire groups (i.e. groups of brothers). We found that, under field conditions, workers from different sire groups, forming a patriline within a given colony, indeed differ in their susceptibility to the common intestinal parasite, Crithidia bombi, and do so independently of queen mating frequency.