The lack of biological knowledge of many invasive species remains as one of the greatest impediments to their management. Here I detail targeted research into the biology of the yellow crazy ant Anoplolepis gracilipes within northern Australia and detail how such knowledge can be used to improve the management outcomes for this species. I quantified nest location and density in three habitats, worker activity over 24 h, infestation expansion rate, seasonal variation of worker abundance and the timing of production of sexuals. Nests were predominantly (up to 68%) located at the bases of large trees, indicating that search efforts should focus around tree bases. Nest density was one nest per 22, 7.1 and 6.3 m2 in the three habitats, respectively. These data form the baselines for quantifying treatment efficacy and set sampling densities for post-treatment assessments. Most (60%) nests were underground, predominantly (89%) occurring in an open area rather than underneath a rock or log. Some seasonality was evident for nests within leaf litter, with most (83%) occurring during the ‘wet season’ (October–March). Of the underground nests, most were shallow, with 44% being less than 10 cm deep, and 67% being less than 20 cm deep. Such nest location and density information serves many management purposes, for improving detection, mapping and post-treatment assessments, and also provided strong evidence that carbohydrate supply was a major driver of A. gracilipes populations. Just over half of the nests (56%) contained queens. Of the 62 underground nests containing queens, most queens (80%) were located at the deepest chamber. When queens were present, most often (38%) only one queen was present, the most being 16. Queen number per nest was the lowest in July and August just prior to the emergence of virgin queens in September, with queen numbers then remaining steadily high until April. Nothing is known for any ant species about how the queen number per nest/colony affects treatment efficacy, but further research would no doubt yield important breakthroughs for treating ants. Activity occurred predominantly nocturnally, ceasing during mid-day. These activity data determined the critical threshold above which work must be conducted to be considered reliable, and also suggests that treatments are best applied in the afternoon. Total brood production peaked in February and was the lowest around August and September. These abundance data form the baselines for quantifying treatment efficacy, and may have implications for treatment efficacy. Males were found every month, predominantly between August and November. Queen pupae were found in September. The reproductive timing of sexuals determines the treatment schedule. Targeted, site-specific research such as that described here should be an integral part of any eradication program for invasive species to design knowledge-based treatment protocols and determine assessment benchmarks.