The expanding global trade in herpetofauna has contributed to new infectious disease dynamics and pathways that allow for the rapid spread of pathogens geographically. Improved biosecurity is needed to mitigate adverse biodiversity, economic and human health impacts associated with pathogen transmission through the herpetological trade. However, general lack of knowledge of the pathogen transmission risks associated with the global trade in herpetofauna and public opposition to biosecurity measures are critical obstacles to successfully preventing pathogen transmission. In 2019 we administered a survey to 2,007 members of the public in the United States of America to ascertain their support for interventions to prevent the spread of Batrachochytrium dendrobatidis (Bd), Batrachochytrium salamandrivorans (Bsal), ranaviruses, and Salmonella through the herpetological trade. We presented survey respondents with different potential hazards associated with pathogen transmission through this trade, namely ecological, economic, and human health impacts. We used structural equation models to determine how these different hazards and respondents’ characteristics influenced respondents’ support for quarantine and veterinary observation of herpetofauna imported into the United States, mandatory tests for diseases of concern, and best practices to reduce stress and improve the care of live herpetofauna during transport to the United States. Respondents’ values and their perceived susceptibility and sensitivity to different hazards associated with pathogen transmission were key determinants of their support for biosecurity. Respondents with strong biospheric and altruistic values demonstrated sensitivity to ecological and human health impacts associated with pathogen transmission, whereas respondents with strong egoistic values demonstrated sensitivity to economic impacts. Respondents had limited knowledge of Bd, Bsal or ranaviruses, the size of the herpetological trade, or how this trade may contribute to pathogen transmission. Improved outreach and education on pathogen transmission through the herpetological trade is required, but it is important that messages are tailored to people with different values to elicit their support for biosecurity.