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Journal of Clinical Microbiology, February 1998, p. 402-408, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Effects of Decontamination Methods and Culture Conditions on Viability of Mycobacterium ulcerans in the BACTEC System

J. C. Palomino* and F. Portaels

Mycobacteriology Unit, Department of Microbiology, Institute of Tropical Medicine Prince Leopold, B-2000 Belgium

Received 29 July 1997/Returned for modification 26 September 1997/Accepted 14 November 1997

We used the BACTEC system to evaluate the effects of several decontamination methods and the addition of antibiotics on the viability of Mycobacterium ulcerans. The effects of polyoxyethylene stearate or egg yolk as supplements were also evaluated to determine their impact on the growth of M. ulcerans. Strains of different geographic origins were subjected to Petroff, reversed Petroff, oxalic acid, and mild HCl treatments. After treatment, the viability of each strain was assessed in the BACTEC system. All of the decontamination methods tested adversely affected bacterial viability. Treatment with mild HCl gave the best results, allowing better growth rates with some strains and causing a delay in growth with others, depending on the geographic origin of the strain. A mixture of polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin did not significantly inhibit growth. Supplementing BACTEC medium with egg yolk markedly improved the recovery of M. ulcerans following the use of each of the decontamination methods. Our findings demonstrate a detrimental impact on the viability of M. ulcerans by all of the decontamination methods currently in common use. This explains, at least in part, the difficulty often experienced in cultivating this organism from clinical specimens. Egg yolk should be added to enhance the rate of successful primary cultivation of M. ulcerans in the BACTEC system.

* Corresponding author. Mailing address: Mycobacteriology Unit, Department of Microbiology, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Belgium. Phone: 32-3 247 6334. Fax: 32-3 247 6333. E-mail: palomino{at}microbiol.itg.be.

Journal of Clinical Microbiology, February 1998, p. 402-408, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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