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Applied and Environmental Microbiology, April 2004, p. 2161-2171, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2161-2171.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Characterization of Bacillus Probiotics Available for Human Use

Le H. Duc,1 Huynh A. Hong,1 Teresa M. Barbosa,2 Adriano O. Henriques,2 and Simon M. Cutting1*

School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 OEX, United Kingdom,1 Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2781-901 Oeiras Codex, Portugal2

Received 24 July 2003/ Accepted 20 December 2003

Bacillus species (Bacillus cereus, Bacillus clausii, Bacillus pumilus) carried in five commercial probiotic products consisting of bacterial spores were characterized for potential attributes (colonization, immunostimulation, and antimicrobial activity) that could account for their claimed probiotic properties. Three B. cereus strains were shown to persist in the mouse gastrointestinal tract for up to 18 days postadministration, demonstrating that these organisms have some ability to colonize. Spores of one B. cereus strain were extremely sensitive to simulated gastric conditions and simulated intestinal fluids. Spores of all strains were immunogenic when they were given orally to mice, but the B. pumilus strain was found to generate particularly high anti-spore immunoglobulin G titers. Spores of B. pumilus and of a laboratory strain of B. subtilis were found to induce the proinflammatory cytokine interleukin-6 in a cultured macrophage cell line, and in vivo, spores of B. pumilus and B. subtilis induced the proinflammatory cytokine tumor necrosis factor alpha and the Th1 cytokine gamma interferon. The B. pumilus strain and one B. cereus strain (B. cereus var. vietnami) were found to produce a bacteriocin-like activity against other Bacillus species. The results that provided evidence of colonization, immunostimulation, and antimicrobial activity support the hypothesis that the organisms have a potential probiotic effect. However, the three B. cereus strains were also found to produce the Hbl and Nhe enterotoxins, which makes them unsafe for human use.

* Corresponding author. Mailing address: School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 OEX, United Kingdom. Phone: 44 1784 443760. Fax: 44 1784 434326. E-mail: s.cutting{at}rhul.ac.uk.

Applied and Environmental Microbiology, April 2004, p. 2161-2171, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2161-2171.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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