The globally invasive Argentine ant (Linepithema humile) possesses a venom lethal to some amphibian species in the invaded range. To test the novel weapons hypothesis (NWH), the effects of the toxin on the cohabiting amphibian species in the ant’s native range need to be investigated. The invader should benefit from the novel chemical in the invaded range, because the species are not adapted, but the venom should not be effective in the native range. We explore the venom effects on juveniles of three amphibian species with different degrees of myrmecophagy inhabiting the ant’s native range: Rhinella arenarum, Odontophrynus americanus, and Boana pulchella. We exposed the amphibians to the ant venom, determined the toxic dose, and evaluated the short- (10 min to 24 h) and medium-term (14 days) effects. All amphibian species were affected by the venom independently of myrmecophagy. In addition to amphibian sensitivity, we discuss how the differential Argentine ant abundance and density in the two ranges could be the key to the susceptibility of amphibians to the venom, resulting in the possibility of NWH. Our results confirm the potential magnitude of the impact of the Argentine ant in successfully invaded areas for the conservation of already threatened amphibians.