An indirect fluorescent antibody test was used to serologically survey Greyhounds from 10 kennels that are part of the racing Greyhound industry in Florida. Age of dogs ranged from 11 months to 11 years. Additionally, 50 adult non-Greyhound pet dogs were consecutively surveyed. Of 393 Greyhounds tested, 181 (46%) were seropositive for babesiosis; pet dogs were seronegative. Slightly higher percentage of seropositive males than females was observed, but this difference was only significant (P less than 0.01) in the 2- to 5-year age class. Male dogs less than 2 years old had significantly (P less than 0.01) lower seroprevalence than did male dogs greater than 2 years old. All 46 Greyhounds that were actively racing at the time of sample collection were seronegative. Dogs were classified into 2 groups on the basis of whether the kennel owner had sought veterinary attention for anemic pups. The 5 kennel owners that had sought veterinary attention (group A) had significantly (P less than 0.01) higher seroprevalence (78.5%), compared with the 5 that had not sought veterinary attention (group B; 23.0%). Seroprevalence of babesiosis in Greyhounds in Florida was comparable to that reported in a limited survey of other southeastern states. It appears to be higher than that in the pet population. Breeding kennels in Florida and other southeastern states from which anemic pups originate should be screened for babesiosis.