Experimental transmission of Hepatozoon americanum to dogs was attempted with four ixodid ticks, viz., Rhipicephalus sanguineus, Amblyomma americanum, Amblyomma maculatum and Dermacentor variabilis. Ticks that dogs ingested included some that were laboratory-reared and experimentally fed as nymphs on a dog with naturally occurring hepatozoonosis; other ticks were collected as replete or partially engorged larvae, nymphs and adults from dogs that had hepatozoonosis and natural infestations of ticks. Whole ticks used to expose susceptible dogs orally were partially dissected to help release oocysts. Among eight dogs exposed, only the three that were fed A. maculatum adults experimentally acquisition fed as nymphs became infected. Dogs developed elevated body temperature and other evidence of clinical disease starting 4 weeks after exposure. 'Cysts' typical of H. americanum were found in skeletal muscle when samples were first examined 5 weeks after dogs ingested ticks, and parasites were also observed in peripheral blood smears at approximately the same time. Our study demonstrates that A. maculatum nymphs can acquire H. americanum by feeding on a parasitemic dog and that transstadial transmission of the protozoan occurs, with dogs acquiring infection when they ingest newly molted adult ticks.