Horned lizards, Phrynosoma spp., and harvester ants, Pogonomyrmex spp., could be in a predator–prey arms race in which the lizards are specialists that feed on harvester ants, and ants have highly toxic venom and other defenses to help deter predacious horned lizards. All 23 examined species of harvester ants possess venoms that are highly lethal to mice, but the venoms of the tested ant species were nearly inactive toward horned lizards. Blood plasma of Phrynosoma cornutum contains a factor (or factors) that neutralizes the ability of harvester ant venom to kill mice, but does not neutralize the venoms of honeybees, a rattlesnake, Russell's viper, or a cobra. A species of harvester ant present only in southern South America was used to test the predictions that the lethality of harvester ant venom evolved in response to predation pressure from horned lizards, and that horned lizard plasma does not neutralize the lethality of this species of harvester ant. This ant species did not overlap in range with horned lizards, which have a range from Guatemala to Canada. Not only was the venom of the South American ant species the most lethal of all tested harvester ant venoms, the venom's lethal activity was neutralized by horned lizard plasma. These results indicate that horned lizards responded to the lethality of their invertebrate prey's venom, but that the harvester ant venom lethality did not evolve in response to predation pressures by present day horned lizards.