To better understand the role of plasmids and their importance in the endemic antibiotic resistance of Enterobacteriaceae, we began a prospective study of our combined medical intensive care/coronary care unit. An initial culture survey of the patients, ward staff, and environment was followed by a prospective sampling of 139 consecutive new admissions at the time of admission to the unit, and at regular intervals thereafter for the remainder of their stay. All cultures were planted on agar-containing gentamicin. Of the 147 patients studied, 12 (8.2%) were colonized with 20 strains of gentamicin-resistant gram-negative bacilli (GRGNB) at 29 sites. An additional four GRGNB were isolated from the environment. Of the 24 GRGNB strains, 7 (29%) Enterobacteriaceae carried plasmids shown to carry the gentamicin-resistance determinant. Plasmids were further characterized by restriction endonuclease digestion profiles of plasmid DNA purified from E. coli C600 transconjugants or transformants. A 93 kb plasmid introduced to the unit by a Serratia liquefaciens colonizing a patient transferred from another area in the hospital was identical to 93 kb plasmids carried by a C. amalonaticus and an E. aerogenes subsequently colonizing another patient on the unit. A 60 kb plasmid, first isolated from a S. marcescens colonizing a sink drain was later isolated from an E. cloacae colonizing a patient. Our results indicate that spread of specific R-plasmids may be one mechanism for dissemination of antibiotic resistance on our MICU/CCU.