Cytauxzoon felis is a long-recognized hemoparasite of free-ranging Florida panthers (Puma concolor coryi), but its prevalence and effect on the population has not been assessed. Red blood cell indices and white blood cell counts were compared between infected and noninfected Florida panthers and Texas cougars (Puma concolor stanleyana) from 1983–1997 in Florida (USA). The prevalence of cytauxzoonosis for both populations was 39% (11/28) for Texas cougars, 35% for Florida panthers (22/63) and 36% overall. Thirteen hematologic parameters were compared between C. felis positive and negative panthers and cougars. Florida panthers had significantly lower mean cell hemoglobin count (MCHC) and higher white blood cell (WBC), neutrophil, monocyte and eosinophil counts (P ≤ 0.05) than Texas cougars. Infected Florida panthers had significantly lower mean cell hemoglobin (MCH) and monocyte counts and higher neutrophil and eosinophil counts than infected Texas cougars. Although statistically significant differences were measured for hematologic parameters in C. felis positive panthers and cougars, biologically significant differences were not likely because values were generally within expected reference ranges for healthy animals. Cytauxzoonosis does not appear to have a negative effect on the hematologic parameters of chronically infected panthers and cougars. Potential transient changes during initial infection were not evaluated.