Aerobic gram-negative commensal faecal flora from 362 healthy volunteers was examined for resistance to trimethoprim. Three hundred fifty-seven trimethoprim-resistant organisms were isolated from 272 of the volunteers (297 Escherichia coli, 46 Klebsiella spp., 9 Enterobacter spp. and 7 other species). Trimethoprim resistance was associated with resistance to other antibiotics at the following frequencies: ampicillin 71.4%, tetracycline 88%, cephalosporins 14% and aminoglycosides 4%. High-level resistance to trimethoprim (MIC > or = 1024 mg/l) occurred in 98.6% of the isolates. Trimethoprim resistance was transferable in 51.2% of the isolates. An X+ factor was required to mobilize resistance in a further 3.4%. Resistance to other antibiotics cotransferred with trimethoprim at the following frequencies: ampicillin 55.4%, tetracycline 30%, cephalosporins 1.5% and aminoglycosides 2.6%. Restriction enzyme analysis of 148 plasmids revealed 79 different profiles. Two restriction profiles represented 10.1 and 8.8% of these plasmids, respectively. The large number of different antibiograms and restriction profiles indicates that there is a large gene pool of trimethoprim-resistant organisms in the faecal flora.