Cane toads are an invasive pest species in which all life stages employ cardiotoxic bufagenins as a chemical defense against predators. Curiously, the bufagenin profiles of eggs and tadpoles are more complex than those of parotoid secretion, the principle mechanism of toxin delivery in adult toads. In an effort to understand this complexity, we determined that selected strains of adult toad parotoid-gland-associated Gram-positive bacteria, Bacillus spp., were capable of biotransforming secreted bufagenins, marinobufagenin (1), telocinobufagenin (2), bufalin (3), and resibufagenin (4), to hydroxylated scaffolds commonly encountered in cane toad eggs and tadpoles. Scaled-up cultivation, preparative chromatography, and detailed spectroscopic analysis identified Bacillus sp. CMB-TD29 biotransformation products of 1, as 11α-hydroxymarinobufagenin (6), 12β-hydroxymarinobufagenin (7), and 17α-hydroxymarinobufagenin (8). Comparative bufagenin profiles across the cane toad life cycle suggest that bacterial biotransformation mediates the oxidative adaptation of adult toad bufagenins to hydroxylated bufagenins encountered in eggs and tadpoles. We speculate that knowledge of a relationship between the cane toad microbiome and bufagenin chemical defenses could inspire the development of a natural, nontoxic, environmentally sustainable bacterial biocontrol for this toxic invasive species.