Many pathogens infect multiple hosts, and spillover from domestic to wild species poses a significant risk for spread of diseases that threaten wildlife and humans. Documentation of cross-species transmission, and unravelling the mechanisms that drive it, remains a challenge. Focusing on co-occurring domestic and wild felids, we evaluate possible transmission mechanisms and evidence of spillover of ‘Candidatus Mycoplasma haemominutum’ (CMhm), an erythrocytic bacterial parasite of cats. We examine transmission and possibility of spillover by analysing CMhm prevalence, modeling possible transmission pathways, deducing genotypes of CMhm pathogens infecting felid hosts based on sequences of the bacterial 16S rRNA gene, and conducting phylogenetic analyses with ancestral state reconstruction to identify likely cross-species transmission events. Model selection analyses suggest both indirect (i.e., spread via vectors) and direct (i.e., via inter-specific predation) pathways may play a role in CMhm transmission. Phylogenetic analyses indicate that transmission of CMhm appears to predominate within host species, with occasional spillover, at unknown frequency, between species. These analyses are consistent with transmission by predation of smaller cats by larger species, with subsequent within-species persistence after spillover. Our results implicate domestic cats as a source of global dispersal and spillover to wild felids via predation. We contribute to the emerging documentation of predation as a common means of pathogen spillover from domestic to wild cats, including pathogens of global conservation significance. These findings suggest risks for top predators as bioaccumulators of pathogens from subordinate species.