A data file consisting of 40 biochemical and physiological tests for the differentiation of aeromonads was constructed based upon existing published data from various geographical locations and laboratories. The data file covers all the current genomospecies (or hybridisation groups) of Aeromonas and is therefore applicable to bacteriologists from a wide variety of fields. The protocol derived from the data file was adapted for use with a semi-interactive computer identification program. This program was challenged with the type strains of all the currently recognised genomospecies and each genomospecies was correctly identified, with only one strain (ATCC 7966, HG1) having an identification probability of less than 90%. The program was also tested with environmental and clinical isolates and again identifications were achieved with high probabilities. This protocol is designed for use on presumptive isolates of Aeromonas spp. and can be carried out in routine laboratories not equipped to perform the complex DNA hybridisation techniques which are currently used for the differentiation of genomospecies of Aeromonas.