The past few decades have seen the emergence of several diseases with drastic public health and economic consequences. Understanding routes of pathogen emergence and transmission is critical to preventing and mitigating disease spillover and amplification. The National Academies Forum on Microbial Threats hosted a public workshop to address gaps in understanding of disease emergence, with a focus on the human-animal interface and laboratory biosafety. Workshop participants explored how applications of existing policy structures, emerging technologies, and actionable research can improve biosecurity measures and prevention of future disease emergence.