Three groups of beagle dogs were fed either a control diet, a diet containing virginiamycin (55 microgram/g of diet), or a diet containing penicillin (110 microgram/g of diet). The proportions of lactose-fermenting organisms in their feces that were resistant to ampicillin, dihydrostreptomycin, tetracycline, or chloramphenicol were measured by a comparative plate-counting procedure. Both antibiotic-supplemented diets resulted in an increase (P < 0.001) in the occurrence of ampicillin, dihydrostreptomycin, and tetracycline resistances during the time of their administration. The occurrence of these resistances was greater (P < 0.001) in the group receiving penicillin than in the group receiving virginiamycin. In addition to the above resistances, a greater (P < 0.001) occurrence of resistance to a sulfonamide (sulfamethoxypyridazine) due to treatment was found by susceptibility testing of isolates. Representative isolates were able to transfer their resistance to a strain of Escherichia coli K-12.