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Applied and Environmental Microbiology, October 1999, p. 4637-4645, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Toxigenic Strains of Bacillus licheniformis Related to Food Poisoning

M. S. Salkinoja-Salonen,1,* R. Vuorio,1 M. A. Andersson,1 P. Kämpfer,2 M. C. Andersson,3 T. Honkanen-Buzalski,4 and A. C. Scoging5

Department of Applied Chemistry and Microbiology1 and Animal Reproduction, Department of Clinical Sciences, Saarentaus,3 FIN-00014 University of Helsinki, and Department of Food Microbiology, National Veterinary and Food Research Institute (EELA), 00231 Helsinki,4 Finland; Institut für Angewandte Mikrobiologie, Justus-Liebig Universität, D-35390 Giessen, Germany2; and Food Hygiene Laboratory, Central Public Health Laboratory, Public Health Laboratory Service, London NW9 5HT, United Kingdom5

Received 18 November 1998/Accepted 5 May 1999

Toxin-producing isolates of Bacillus licheniformis were obtained from foods involved in food poisoning incidents, from raw milk, and from industrially produced baby food. The toxin detection method, based on the inhibition of boar spermatozoan motility, has been shown previously to be a sensitive assay for the emetic toxin of Bacillus cereus, cereulide. Cell extracts of the toxigenic B. licheniformis isolates inhibited sperm motility, damaged cell membrane integrity, depleted cellular ATP, and swelled the acrosome, but no mitochondrial damage was observed. The responsible agent from the B. licheniformis isolates was partially purified. It showed physicochemical properties similar to those of cereulide, despite having very different biological activity. The toxic agent was nonproteinaceous; soluble in 50 and 100% methanol; and insensitive to heat, protease, and acid or alkali and of a molecular mass smaller than 10,000 g mol-1. The toxic B. licheniformis isolates inhibited growth of Corynebacterium renale DSM 20688T, but not all inhibitory isolates were sperm toxic. The food poisoning-related isolates were beta-hemolytic, grew anaerobically and at 55°C but not at 10°C, and were nondistinguishable from the type strain of B. licheniformis, DSM 13T, by a broad spectrum of biochemical tests. Ribotyping revealed more diversity; the toxin producers were divided among four ribotypes when cut with PvuII and among six when cut with EcoRI, but many of the ribotypes also contained nontoxigenic isolates. When ribotyped with PvuII, most toxin-producing isolates shared bands at 2.8 ± 0.2, 4.9 ± 0.3, and 11.7 ± 0.5 or 13.1 ± 0.8 kb.

* Corresponding author. Mailing address: Department of Applied Chemistry and Microbiology, P.O. Box 56 (Biocenter), 00014 University of Helsinki, Finland. Phone: 358-9-70859300. Fax: 358-9-70859301. E-mail: mirja.salkinoja-salonen{at}helsinki.fi.

Applied and Environmental Microbiology, October 1999, p. 4637-4645, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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