Hygienic behavior in stingless bees is a trait of workers that confers colony-level resistance against some brood diseases. Workers of hygienic colonies detect, uncap and remove dead or diseased brood from the nest cells. We examined the hygienic behavior in stingless bees (Melipona quadrifasciata anthidioides ) from freezekilled brood assay using liquid nitrogen. Responses were measured at 14 times (3, 6, 12, 24, 48, 72, 96, 120, 144, 168, 192, 216, 240 and 264 h after freeze-killing of the brood). Workers were estimated to remove on average 65% of larvae and 34% of dead pupae within 48 h of freezing. Workers removed dead brood rapidly after uncapping the cells. Strong colonies showed a greater removal of dead pupae, while the size of the population did not influence the removal of dead larvae. These findings report for the first time the hygienic behavior in M. q. anthidioides and confirm that workers have more difficulty removing pupae compared with larvae from the combs.generalized linear models / stingless bees / hygienic colonies / dead brood removal / freeze-killed brood