Environmental factors have been associated with the production of aflatoxin in maize, Zea mays L., and it is inconclusive whether transgenic, Bacillus thuringiensis (Bt), maize has an impact on aflatoxin accumulation. Maize hybrids differing in transgenes were planted in two locations from 2014 to 2017. Yield, aflatoxin, and ear injury caused by corn earworm, Helicoverpa zea (Boddie), and fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), were measured across three groups of hybrids differing in transgenes including near-isogenic hybrids, and water-stressed conditions. The hybrid groups consisted of non-Bt hybrids with no Bt transgenes, a second group with one or more Cry-Bt transgenes, and the third group with vegetative insecticidal Bt protein and Cry-Bt transgenes (Cry/Vip-Bt). Across the six data sets derived from 11 experiments, the Cry-Bt and Cry/Vip-Bt hybrids had less ear injury and aflatoxin on average than non-Bt hybrids. The effects of ear injury on yield and aflatoxin were more prominent and consistent in Corpus Christi, TX, where hybrids experienced more water-limited conditions than in College Station, TX. The trend of increased aflatoxin among hybrids with increased ear injury was further resolved when looking at Cry-Bt and Cry/Vip-Bt isogenic hybrids in Corpus Christi. The results supported that the maize hybrids with the inclusion of Cry-Bt and Cry/Vip-Bt transgenes warrant further investigation in an integrated approach to insect and aflatoxin management in sub-tropical rain-fed maize production regions. Research outcomes may be improved by focusing on areas prone to water-stress and by using hybrids with similar genetic backgrounds.