Veterinary and animal care workers perform critical functions in biosecurity and public health, yet little has been done to understand the unique needs and barriers these workers face when responding during a pandemic crisis. In this article, we evaluate the perceived risks and roles of veterinary and animal care workers during the COVID-19 pandemic, and explored barriers and facilitators in their readiness, ability, and willingness to respond during a pandemic. We deployed a survey targeting US veterinary medical personnel, animal shelter and control workers, zoo and wildlife workers, and other animal care workers. Data were collected on respondents' self-reported job and demographic factors, perceptions of risk and job efficacy, and readiness, ability, and willingness to respond during the pandemic. We found that leadership roles and older age had the strongest association with decreased perceived risk and improved job efficacy and confidence, and that increased reported contact level with others (both coworkers and the public) was associated with increased perceived risk. We determined that older age and serving in leadership positions were associated with improved readiness, willingness, and ability to respond. Veterinary and animal care workers' dedication to public health response, reflected in our findings, will be imperative if more zoonotic vectors of SARS-CoV-2 arise. Response preparedness in veterinary and animal care workers can be improved by targeting younger workers not in leadership roles through support programs that focus on improving job efficacy and confidence in safety protocols. These findings can be used to target intervention and training efforts to support the most vulnerable within this critical, yet often overlooked, workforce.