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Phenotypic consequences of red–white colony type variation in Mycobacterium avium

Seattle Biomedical Research Institute, 4 Nickerson Street, WA 98109, Seattle, USA¹
Kuzell Institute of Arthritis and Infectious Disease, San Francisco, CA, USA²

Author for correspondence: Gerard A. Cangelosi. Tel: +1 206 284 8846. Fax: +1 206 284 0313. e-mail: gcang{at}sbri.org

Mycobacterium avium undergoes reversible morphotypic switching between the virulent transparent colony type and the less virulent opaque colony type. A new morphotypic switch in M. avium, termed red–white, that becomes visible when opaque colonies of clinical isolates are grown on agar media containing Congo red, was recently described. White opaque (WO) variants were found to be more resistant to multiple antibiotics than were red opaque (RO) variants. The present paper reports that transparent derivatives of RO and WO clones retain the differential Congo red binding properties of their opaque parents, indicating that the opaque–transparent switch operates independently of the red–white switch. White transparent variants were more resistant to clarithromycin and rifampin in vitro, and better able to survive within human macrophages, than their red transparent counterparts. Neither red nor white variants were markedly favoured during growth in vitro; however, red variants were better able to spread on soft agar (sliding motility), a potential selective advantage under some environmental circumstances. White-to-red switching was frequently observed in vitro and was accompanied by decreased antibiotic resistance and increased motility. Red-to-white switching has yet to be observed in vitro, indicating that the red morphotype is very stable. Significantly, some widely studied laboratory reference strains of M. avium, including strain 2151 and the genome sequence strain 104, are stable red clones. These strains are intrinsically antibiotic resistant and virulent in animal models, but they may not express genes encoding the elevated levels of antibiotic resistance and intracellular survival observed in white variants.

Keywords: AIDS, drug resistance, virulence, motility, phenotypic switching

Abbreviations: CR, Congo red; DR, dark red; relative Congo red binding; RO, red opaque; RT, red transparent; WO, white opaque; WT, white transparent; ZOI, zone of inhibition

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