Male CD-1 mice, fed streptomycin in their drinking water, were used to study colonization of the mouse intestine by both fecal Escherichia coli strains isolated from healthy humans and Escherichia coli K12 strains which are routinely used as hosts for recombinant DNA. Prior to use in mice, all the strains were made resistant to streptomycin. Several facts emerged from these studies: (a) Strains isolated from different healthy humans colonized the mouse intestine with equal ability (approximately 10(8) cells/g feces), but may have colonized biochemically distinct sites. (b) K12 strains tested had, at most, one hundredth the colonizing ability of human fecal strains. (c) Rifampicin-resistant mutants of strains which contain one or no plasmids were poor colonizers relative to their parents. (d) Rifampicin-resistant mutants of strains which contain six or more plasmids retained the colonizing abilities of their parents. (e) Introduction of the F-amp or pJBK5 plasmid into HS-4, a human fecal strain which does not normally carry these plasmids, reduced its colonizing ability 1000-fold. (f) Strains used in this study colonized the mouse caecum and colon exclusively. The system presented here offers a simple, rapid test to determine whether a specific genetic alteration in a bacterium (e.g. antibiotic resistance) results in enhanced, reduced, or unchanged colonizing ability. Such a test might prove to be of value as a part of the clinical testing of antibiotics.