Amphibians and reptiles are declining globally. One potential cause of this decline includes impacts resulting from co-occurrence with non-native red imported fire ant, Solenopsis invicta. Although a growing body of anecdotal and observational evidence from laboratory experiments supports this hypothesis, there remains a lack of field scale manipulations testing the effect of fire ants on reptile and amphibian communities. We addressed this gap by measuring reptile and amphibian (“herpetofauna”) community response to successful fire ant reductions over the course of 2 years following hydramethylnon application to five 100–200 ha plots in southeastern coastal South Carolina. By assessing changes in relative abundance and species richness of herpetofauna in response to fire ant reductions, we were able to assess whether some species were particularly vulnerable to fire ant presence, and whether this sensitivity manifested at the community level. We found that herpetofauna abundance and species richness responded positively to fire ant reductions. Our results document that even moderate populations of red imported fire ants decrease both the abundance and diversity of herpetofauna. Given global herpetofauna population declines and continued spread of fire ants, there is urgency to understand the impacts of fire ants beyond anecdotal and singles species studies. Our results provides the first community level investigation addressing these dynamics, by manipulating fire ant abundance to reveal a response in herpetofauna species abundance and richness.