A total of 270 recent nosocomial blood stream isolates of non-Enterobacteriaceae Gram-negative bacilli representing nearly 50 U.S. medical centers were characterized. The numbers of isolates of individual organisms were: Pseudomonas aeruginosa (n = 204), Acinetobacter spp. (n = 48), aid Stenotrophomonas maltophilia (n = 18). MICs were determined using the broth microdilution susceptibility method with 12 antimicrobial agents: piperacillin, piperacillin/tazobactam, ceftriaxone, ceftazidime, cefepime, imipenem, ciprofloxacin, ofloxacin, amikacin, gentamicin, tobramycin, and tri methoprim/sulfamethoxazole. Based on current National Committee far Clinical Laboratory Standards breakpoints, rates of resistance to cefepime, ceftazidime, and imipenem were as follows : P. aeruginosa, 3, 9, and 5%; Acinetobacter spp., 2, 37, and 0%; and S. maltophilia, 88.7, 35.3, and 100%, respectively. Trimethoprim/sulfamethoxazole was the most active agent against S. maltophilia (100% susceptible). Twenty-eight isolates of P. aeruginosa that expressed high levels of resistance to ceftazidime (MIC, >256 mu g/mL) and imipenem (MIG, >32 mu g/mL) were examined for potential metallo-beta-lactamase production by polymerase chain reaction and were found to be negative. Molecular typing of P. aeruginosa isolates revealed many patient-unique strains, but also noted clustering of infections due to isolates of the same DNA type, suggesting possible nosocomial transmission in 9 of 14 medical centers. Given the resistance profile and pathogenic potential of these non-enteric Gram-negative bacilli, considerable effort should be exerted to develop and enforce infection control and antimicrobial utilization practices that will limit the spread of these organisms in the hospital environment. (C) 1997 Elsevier Science Inc.