Abstract In phylogeography, ancestral state inference methods are used to identify the geographic or host species origin of viral or bacterial lineages and reconstruct their transmission histories over time. However, differences in sampling among states can bias these inference methods. Here, we introduce sampling-aware ancestral state inference (SAASI), a method that accounts for sampling differences. We apply SAASI to the multi-host spread of the H5N1 virus in the United States in 2024 and find that the key transmission event from wild birds to cattle is estimated to occur later under lower sampling in wild birds (compared to other species) than when sampling is not accounted for. Using simulation, we find that SAASI infers past viral locations/host species considerably more accurately than standard methods when sampling bias exists, is computationally feasible for large datasets, and scales to trees with 100,000 tips.
Song et al. (2026) studied this question.