Successful Cytauxzoon felis transmission studies have occurred using Amblyomma americanum adults acquisition-fed as nymphs on an experimentally infected domestic cat or Dermacentor variabilis adults fed as nymphs on a splenectomized bobcat. Here, we evaluated A. americanum and D. variabilis nymphs acquisition-fed as larvae on a C. felis-infected carrier domestic cat for competence to transmit the protozoan parasite as nymphs to naïve, healthy domestic cats.Amblyomma americanum and D. variabilis larvae were applied to a chronically infected, parasitemic C. felis donor cat (Felis catus) and allowed to feed to repletion. Engorged larvae were collected and held through ecdysis. Three cats were each infested with 66 A. americanum or 66 D. variabilis emerged nymphs. Cytauxzoon felis infections in principal cats were determined by clinical signs and detection of circulating parasite by blood smear and PCR evaluation.Clinical signs of cytauxzoonosis were observed in cats infested with A. americanum nymphs beginning 12-15 days post-infestation (dpi). The same cats were PCR positive on 12-14 dpi; piroplasms were evident in blood smears at 16 dpi, and macrophage schizonts were observed in stained spleen impression smears in two animals at necropsy. Cats infested with acquisition-fed D. variabilis nymphs remained clinically normal and did not develop detectable parasitemia over the course of the study as determined by blood smear and PCR.Cytauxzoon felis was successfully transmitted to domestic cats by A. americanum nymphs acquisition-fed as larvae on the donor cat. However, we were not able to transmit C. felis to healthy domestic cats with D. variabilis nymphs that were simultaneously acquisition-fed on the same donor cat. Results from this study suggest that larval and nymphal A. americanum likely play important roles in natural transmission cycles of C. felis.