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Antimicrobial Agents and Chemotherapy, February 2000, p. 294-303, Vol. 44, No. 2
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of Passage-Selected Vancomycin-Resistant Staphylococcus aureus Strains of Diverse Parental Backgrounds

Richard F. Pfeltz, Vineet K. Singh, Jennifer L. Schmidt, Michael A. Batten, Christopher S. Baranyk, Matthew J. Nadakavukaren, Radheshyam K. Jayaswal, and Brian J. Wilkinson*

Microbiology Group, Department of Biological Sciences, Illinois State University, Normal, Illinois 61790-4120

Received 5 April 1999/Returned for modification 27 June 1999/Accepted 25 October 1999

A series of 12 Staphylococcus aureus strains of various genetic backgrounds, methicillin resistance levels, and autolytic activities were subjected to selection for the glycopeptide-intermediate S. aureus (GISA) susceptibility phenotype on increasing concentrations of vancomycin. Six strains acquired the phenotype rapidly, two did so slowly, and four failed to do so. The vancomycin MICs for the GISA strains ranged from 4 to 16 µg/ml, were stable to 20 nonselective passages, and expressed resistance homogeneously. Neither ease of acquisition of the GISA phenotype nor the MIC attained correlated with methicillin resistance hetero- versus homogeneity or autolytic deficiency or sufficiency. Oxacillin MICs were generally unchanged between parent and GISA strains, although the mec members of both isogenic methicillin-susceptible and methicillin-resistant pairs acquired the GISA phenotype more rapidly and to higher MICs than did their susceptible counterparts. Transmission electron microscopy revealed that the GISA strains appeared normal in the absence of vancomycin but had thickened and diffuse cell walls when grown with vancomycin at one-half the MIC. Common features among GISAs were reduced doubling times, decreased lysostaphin susceptibilities, and reduced whole-cell and zymographic autolytic activities in the absence of vancomycin. This, with surface hydrophobicity differences, indicated that even in the absence of vancomycin the GISA cell walls differed from those of the parents. Autolytic activities were further reduced by the inclusion of vancomycin in whole-cell and zymographic studies. The six least vancomycin-susceptible GISA strains exhibited an increased capacity to remove vancomycin from the medium versus their parent lines. This study suggests that while some elements of the GISA phenotype are strain specific, many are common to the phenotype although their expression is influenced by genetic background. GISA strains with similar glycopeptide MICs may express individual components of the phenotype to different extents.

* Corresponding author. Mailing address: Department of Biological Sciences, Campus Box 4120, Illinois State University, Normal, IL 61790-4120. Phone: (309) 438-7244. Fax: (309) 438-3722. E-mail: bjwilkin{at}ilstu.edu.

Antimicrobial Agents and Chemotherapy, February 2000, p. 294-303, Vol. 44, No. 2
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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