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Journal of Bacteriology, August 2006, p. 5333-5344, Vol. 188, No. 15
0021-9193/06/$08.00+0     doi:10.1128/JB.00303-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Characterization of Bacillus anthracis-Like Bacteria Isolated from Wild Great Apes from Côte d'Ivoire and Cameroon

Silke R. Klee,^1* Muhsin Özel,¹ Bernd Appel,^1, Christophe Boesch,² Heinz Ellerbrok,¹ Daniela Jacob,¹ Gudrun Holland,¹ Fabian H. Leendertz,^1,2,3 Georg Pauli,¹ Roland Grunow,¹ and Herbert Nattermann¹

Robert Koch Institut, Centre for Biological Safety, Berlin, Germany,¹ Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany,² Great Ape Health Monitoring Unit, Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany³

Received 1 March 2006/ Accepted 12 May 2006

We present the microbiological and molecular characterization of bacteria isolated from four chimpanzees and one gorilla thought to have died of an anthrax-like disease in Côte d'Ivoire and Cameroon. These isolates differed significantly from classic Bacillus anthracis by the following criteria: motility, resistance to the gamma phage, and, for isolates from Cameroon, resistance to penicillin G. A capsule was expressed not only after induction by CO₂ and bicarbonate but also under normal growth conditions. Subcultivation resulted in beta-hemolytic activity and gamma phage susceptibility in some subclones, suggesting differences in gene regulation compared to classic B. anthracis. The isolates from Côte d'Ivoire and Cameroon showed slight differences in their biochemical characteristics and MICs of different antibiotics but were identical in all molecular features and sequences analyzed. PCR and Southern blot analyses confirmed the presence of both the toxin and the capsule plasmid, with sizes corresponding to the B. anthracis virulence plasmids pXO1 and pXO2. Protective antigen was expressed and secreted into the culture supernatant. The isolates possessed variants of the Ba813 marker and the SG-749 fragment differing from that of classic B. anthracis strains. Multilocus sequence typing revealed a close relationship of our atypical isolates with both classic B. anthracis strains and two uncommonly virulent Bacillus cereus and Bacillus thuringiensis isolates. We propose that the newly discovered atypical B. anthracis strains share a common ancestor with classic B. anthracis or that they emerged recently by transfer of the B. anthracis plasmids to a strain of the B. cereus group.

Present address: Federal Institute for Risk Assessment, Berlin, Germany.

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