Abstract Disease-associated mortality is a leading cause of amphibian declines and extinctions worldwide. Understanding the influence of land-management practices, like herbicide use, on amphibian immune defense traits could guide changes to improve conservation outcomes. Amphibians are partially protected from pathogens by two skin-associated immune defense traits: bacterial communities inhabiting their skin, and antimicrobial peptides secreted by the skin. Utilizing the Blanchard's Cricket Frog (Acris blanchardi), a declining North American amphibian species, as our model, we manipulated Rodeo™ aquatic herbicide concentration and the life stage at which Rodeo exposure occurred. We assessed juvenile survival, time to metamorphosis, juvenile mass, and skin-associated immune defense traits. We found a 37% decrease in survival of larvae exposed to 2.5 mg a.e. L−1 (acid equivalent) compared to controls despite that this commercial herbicide formulation does not contain an added surfactant. Surviving larvae e...