Fitness Cost Due to Mutations in the 16S rRNA Associated with ...

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Jun 22, 2005 - The fitness cost of a resistance determinant is the primary parameter that determines its frequency in vivo. ... Mailing address: Department of Micro- ...... ments as a consequence of recurrent infections, repeated mass.
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Nov. 2005, p. 4455–4464 0066-4804/05/$08.00⫹0 doi:10.1128/AAC.49.11.4455–4464.2005 Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Vol. 49, No. 11

Fitness Cost Due to Mutations in the 16S rRNA Associated with Spectinomycin Resistance in Chlamydia psittaci 6BC Rachel Binet and Anthony T. Maurelli* Department of Microbiology and Immunology, F. Edward He´bert School of Medicine, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, Maryland 20814-4799 Received 22 June 2005/Returned for modification 26 July 2005/Accepted 2 August 2005

The fitness cost of a resistance determinant is the primary parameter that determines its frequency in vivo. As a model for analysis of the impact of drug resistance mutations on the intracellular life cycle of Chlamydia spp., we studied the growth of four genetically defined spectinomycin-resistant (Spcr) clonal variants of Chlamydia psittaci 6BC isolated in the plaque assay. The development of each variant was monitored over 46 h postinfection in the absence of drug, either in pure culture or in 1:1 competition with the parent strain. Spcr mutations in the 16S rRNA gene at positions 1191 and 1193 were associated with a marked impairment of C. psittaci biological fitness, and the bacteria were severely outcompeted by the wild-type parent. In contrast, mutations at position 1192 had minor effects on the bacterial life cycle, allowing the resistant isolates to compete more efficiently with the wild-type strain. Thus, mutations with a wide range of fitness costs can be selected in the plaque assay, providing a new strategy for prediction and monitoring of the emergence of antibiotic resistance in chlamydiae. So far, drug resistance has not been a serious threat for the treatment of chlamydial infections. Tetracycline is an effective antichlamydial drug that targets 16S rRNA. Attempts to isolate spontaneous tetracycline-resistant mutants of C. psittaci 6BC revealed a frequency