The Biolog Identification System (Biolog, Inc., Hayward, Calif.) is a new bacterial identification method that establishes an identification based on the exchange of electrons generated during respiration, leading to a subsequent tetrazolium-based color change. This system tests the ability of a microorganism to oxidize a panel of 95 different carbon sources. We report on a preliminary investigation of the ability of the instrument to identify, using its computer-driven enzyme immunoassay reader, a diverse group of clinically relevant members of the family Enterobacteriaceae and gram-negative non-Enterobacteriaceae. The Biolog reported identifications (correct or incorrect) for 266 of 352 organisms tested (75.6%). Of the 266 identifications reported, 87.3% were correct at the genus level and 75.6% were correct at the species level at 24 h. In the total study of 352 strains, 46.6% were correct to the species level at 4 h and 57.1% were correct to the species level at 24 h. The error rate was 10.4% after 4 h and 9.6% after 24 h. The Biolog performed well with many genera, but problems were encountered with some strains of Klebsiella, Enterobacter, and Serratia. We found the system to be versatile and easy to use.