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Because West Nile (WN) virus primarily cycles between mosquitoes and birds, North American reptiles and amphibians have not been evaluated as reservoir hosts of this virus. We infected three species of reptiles and one species of amphibian: Iguana iguana (green iguana), Thamnophis sirtalis sirtalis (Florida garter snake), Trachymes scripta elegans (red-ear slider), and Rana catesbeiana (North American bullfrog). After inoculation with WN virus, some of the green iguanas, bullfrogs, and garter snakes showed low but detectable viral loads in the blood, oral or cloacal swabs, and/or organs.