Nepotism shapes interactions among the members of almost every animal society. However, clear evidence of nepotism within highly cooperative insect societies, such as ant, wasp and honeybee colonies, is rare. Recent empirical findings suggest that nepotism occurs within honeybee colonies where kin-selection theory most strongly predicts its existence: during the lethal queen-queen duels that determine which of several young queens will become the colony's next queen. In this study, I test whether worker bees act nepotistically by hindering duelling queens that are distantly related to themselves. I accomplished this by observing labelled workers harassing duelling queen bees in observation hives and subsequently by determining worker-queen relatedness using DNA microsatellites. I show that the workers that harassed duelling queens were neither more-closely nor more-distantly related to them than were workers selected randomly from the colony. Thus, workers did not behave nepotistically by hindering half-sister queens more than full-sister queens. These results demonstrate that under certain conditions, natural selection limits the evolution of nepotism within animal societies despite strong theoretical predictions for its existence.