Simple nonlethal blood culture methodology, an alternative to euthanasia for diagnosing systemic bacterial infections in fish, is described. Blood was extracted from the caudal vein of 20 individuals of five fish species, incubated in brain–heart infusion broth, and then plated onto enriched blood agar. Nine of these fish were subsequently euthanized and necropsied for confirmatory tissue cultures. Five species of bacteria were isolated from the blood cultures from nine fish, and the tissue culture results in euthanized, necropsied fish agreed with the blood culture results in all cases. All the fish that were not euthanized survived for 24 hr, although two heavily parasitized fish subsequently died.