Significance Emerging disease epidemics often result from a pathogen establishing transmission in a novel host species. However, for reasons that remain poorly understood, most cross-species transmissions fail to establish in the newly infected species. Examining experimental cross-species inoculations of rabies virus, we show that host and viral factors predict differences in disease progression in ways that are expected to impact the likelihood of onward transmission. Disease progression was accelerated and virus excretion decreased when the reservoir and novel host were physiologically or genetically more dissimilar. These insights may help to explain and predict host shifts in rabies and other zoonotic viruses and highlight meta-analyses of experimental inoculation data as a powerful and generalizable approach for understanding the dynamics of index infections.