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The secondary succession of tropical rainforest in abandoned agricultural fields modifies components of species diversity and processes of species colonization and replacement. In general, knowledge on invertebrate reassembly is lacking, even though invertebrate assemblages directly influence the maturation of forests. Ants are especially useful for testing hypotheses about the effects of resource diversity and microhabitat conditions. We experimentally assessed the effects of different successional stages on nest colonization. Then, we assessed whether nesting resources and microhabitat conditions (i.e., abiotic conditions) are potential mechanisms that influence the colonization pattern of twig-dwelling ants along a natural chronosequence for 1 year in the Lacandon region, Mexico. We found that ant species richness in twigs is correlated with total species found in leaf litter. The nest occupancy increased as succession progressed, but species richness did not change. Nests occupation increased to greater artificial nesting resource decomposition rates and lower soil pH. Meanwhile, species richness was increased only with lower soil compaction. Species composition changes among successional categories; the changes were greater with greater soil compaction and soil pH. The diversity of nesting resources did not drive twig-dwelling ant assembly. We conclude that the natural maturation of tropical forest enables greater colonization of twig-dwelling ants, but recovery of the species composition of assemblages after 30 years is incomplete, although it shows a recovery trend.