Twelve hundred enterobacterial Escherichia coli isolates of porcine origin were screened phenotypically for antibiotic resistance. The bacteria were isolated from 10 herds of swine with different histories of exposure to antimicrobial agents for therapeutic purposes. The bacterial isolates were part of the normal bacterial flora of the intestines of the animals because they were isolated from healthy individuals. The strains were tested for phenotypic antibiotic resistance against sulfonamides, trimethoprim, streptomycin, ampicillin, neomycin, chloramphenicol, and tetracycline. Resistance against streptomycin was found to be most common, followed by resistance against sulfonamides and tetracycline. The highest number of resistant bacteria was found in herds where the use of antimicrobial agents was considered to be high. A selection of multiresistant bacterial isolates were further genetically characterized by hybridization with probes specific for the antibiotic resistance genes; sulI, sulII, dfrI, dfrIIb, dfrIX, and the class A, B, C, and D tetracycline resistance determinants. A PCR was developed and used for detection of the strA-strB gene pair encoding streptomycin resistance in gram-negative bacteria. The strA-strB gene pair was the most frequent resistance determinant in the isolates examined. This study indicates that nonpathogenic E. coli from swine may represent a considerable reservoir of antibiotic resistance genes that might be transferable to pathogens.