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Road-kills are a major cause of mortality for a wide variety of herpetofauna, but management decisions on remediation procedures for reducing losses are based in economic realities. Because funding is finite for species conservation, bioeconomic analysis can assist in justifying, evaluating, and maximizing returns on conservation expenditures, especially for low-profile species such as herpetofauna. Here, we present a bioeconomic analysis of road-killed herpetofauna in Jonathan Dickinson State Park, Florida. Road surveys were conducted daily for four years to identify and enumerate the numbers of each reptile and amphibian species killed by vehicles. Conservative individual valuations applied to the losses formed the basis of a benefit–cost analysis aimed at identifying the thresholds at which remediation expenditures would be justified. We found an average of 64 reptiles and amphibians were killed/year, justifying conservation expenditures up to $32,000/year. However, if less conservative valuations were applied, especially for threatened and endangered species, justifiable expenditures rise dramatically.