Rickettsia felis can be maintained in cat fleas by vertical transmission for up to 12 generations without the benefit of an infected host. Horizontal transmission or the acquisition of R. felis by fleas feeding on cats or artificially infected meals were not demonstrated in this study. Horizontal transmission of R. felis by the ingestion of feces or eggs by flea larvae was not detected. We also tested for and found no evidence to support horizontal transmission by contact among positive fleas and negative fleas.