The nesting behavior of the euglossine beeEulaema nigrita was observed in the laboratory after being transferred from three nests to observation boxes. Nests were re-used by successive generations with more than one female working in each re-use process. Associated females were always of the same generation. All females that shared a nest foraged for construction materials (mud, excrement and resin) and each constructed, provisioned and oviposited in her own cells. The number of cells constructed by each female ranged from one to 23. Females stayed in the nests for periods ranging from 15 to 59 days during the hot and wet season and from five to 78 days during the cool and dry season. The egg-to-adult period was related to climatic conditions, and in all re-use processes it was longer than the time of residence of the females in the nests. The meloid beetleMeloetyphlus attacephalus was the only nest parasite. Due to the attack method employed by this parasite, the presence of more than one female in the nest did not result in improved nest defense.