Neotropical leaf-cutting ants (tribe Attini) live in obligate symbiosis with fungus they culture for food. To protect themselves and their fungus garden from pathogens, they minimize the entry of microorganisms through mechanical and chemical means. In this study, focusing on the species Acromyrmex subterraneus and A. octospinosus, (Hymeoptera: Formicidae). Self- and allo-grooming behavior were quantified and it was found that A. octospinosus workers spend less time in self-grooming than A. subterraneus. In the experimental absence of fungus in A. subterraneus, the times spent in these two behaviors are not affected; however workers spend significantly more time immobile. Hygienic and trophallaxis behaviors were examined as well as the possibility that workers exchange food, and the grooming behavior of foraging and non-foraging workers were compared. Behavioral observations revealed that large workers spent more time grooming than small workers, and more than 62% of replete foragers passed collected liquid food via trophallaxis to a nestmate. However, trophallaxis was rarely observed between non-forager workers. These results suggest that trophallaxis permits the exchange of alimentary liquid between colony members, but it is not important for spreading the colony odor signature.