Mechanisms of high streptomycin resistance (8000 micrograms/ml) in S. derby cells carrying R plasmids were studied. The cells were isolated from clinical materials. The findings showed that the streptomycin resistance determinant in the S. derby cells was localized on the plasmid. In cell-free extracts of the strains, there was detected no inactivation of aminoglycosides by phosphorylation, adenylation and acetylation of the antibiotic molecules. The plasmid elimination from the cells of S. derby K89 by ethidium bromide resulted in loosing of streptomycin resistance by the cells. This indirectly excluded the mechanism associated with modification of the ribosomes. Streptomycin resistance in the strains studied must be due to decreased permeability of the S. derby K89 cell envelopes for streptomycin.